Nitrogen-containing spiro cyclic compounds and pharmaceutical uses thereof

ABSTRACT

A compound of the following general formula [I]:wherein each symbol has the same meaning as defined herein, or a pharmaceutically acceptable salt thereof, or a solvate thereof, and a pharmaceutical use of the same in treating organ transplant rejection, graft versus host reaction after transplantation, autoimmune disease, allergic disease and chronic myeloproliferative disease.

TECHNICAL FIELD

The present invention relates to a novel nitrogen-containing spirocyclic compound and a pharmaceutical use thereof. Specifically, thepresent invention relates to an inhibitor of Janus kinase 3, referred toas JAK3 hereinafter, a compound for prevention or treatment of organtransplant rejection, graft versus host reaction after transplantation,autoimmune disease and allergic disease, and a pharmaceutical use of thesame.

The present invention also relates to an inhibitor of Janus kinase 2,referred to as JAK2 hereinafter, a compound for prevention or treatmentof chronic myeloproliferative disease, and a pharmaceutical use of thesame.

BACKGROUND ART

JAK3 is a member of Janus family which belongs to protein kinases. Othermembers of this family are expressed in various tissues, while JAK3 isexpressed only in hematopoietic cells.

This limiting expression is involved in an important role of JAK3 by anon-covalent association of JAK3 with γ-chains common to multiply-linkedreceptors including IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 in thereceptor-mediated signal transduction.

Significantly reduced JAK3 protein levels or gene defects in commonγ-chains are found in Severe Combined Immunodeficiency, referred to asSCID hereinafter, patient population. That indicates that animmunosuppression is produced by blocking JAK3-mediated signalingpathway.

It has been reported in animal experiments that JAK3 plays an importantrole in maturity of NK cells, B-lymphocytes and T-lymphocytes and isessentially required for the maintenance of T cell functions.

It has been also reported that a JAK3 inhibitor CP-690,550((3R,4R)-3-[4-methyl-3-[N-methyl-N-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino]piperidin-1-yl]-3-oxopropionitrile)improves rheumatoid arthritis and psoriasis conditions, and showsrejection-suppression effects in simian renal transplantation model andairway inflammation-suppression effects in mouse asthma model.

In view of the above knowledge, it is believed that a regulation of animmune activity by JAK3 inhibitors is useful for the prevention ortreatment of organ transplant rejection, graft versus host reactionafter transplantation, autoimmune disease and allergic disease.

On the other hand, it has been indicated that an inhibition of JAK2 isuseful for patients suffering from a chronic myeloproliferative disease.The chronic myeloproliferative disease includes polycythemia vera,primary myelofibrosis, essential thrombocythemia, chronic myelocyticleukemia, chronic myelomonocytic leukemia, chronic eosinophilicleukemia, chronic neutrophilic leukemia, systemic mastocytosis.

It is believed that the chronic myeloproliferative disease may be causedby acquired cell mutations in hematopoietic stem cells, and it has beenreported that a large majority of polycythemia vera's patients as wellas a significant number of primary myelofibrosis's and essentialthrombocythemia's patients have gain-of-function mutations of JAK2. Ithas been also reported that an inhibition of a JAK2V617F kinase bylow-molecular inhibitors causes an inhibition of proliferation ofhematopoietic cells.

In view of the above knowledge, it is believed that a regulation ofproliferation of hematopoietic cells by JAK2 inhibitors is useful forthe prevention or treatment of chronic myeloproliferative diseases.

Four types of members of a Janus kinase, referred to as JAK hereinafter,family are known including Janus kinase 1, referred to as JAK1hereinafter, JAK2, JAK3, and tyrosine kinase 2, referred to as Tyk2hereinafter, and it is believed that a JAK1 inhibitor and a Tyk2inhibitor are also useful for the prevention or treatment of varietiesof diseases similar to a JAK3 inhibitor.

DISCLOSURE OF INVENTION Problems to be Resolved by the Invention

According to extensive studies for the purpose of developing a noveltherapeutic or preventive agent for organ transplant rejection, graftversus host reaction after transplantation, autoimmune disease andallergic disease alternative to conventional one, the inventors havefound novel nitrogen-containing spiro cyclic compounds with JAK3inhibitory effect and achieved the present invention.

The present inventors have also found novel nitrogen-containing spirocyclic compounds with JAK2 inhibitory effect and achieved the presentinvention.

Means of Solving the Problems

Specifically, the present invention is as follows.

[1] A compound of the following general formula [I]:

wherein R^(a) is the same or different and each:(1) C₁₋₆ alkyl, or(2) halogen atom,

n1 is an integer selected from 0 to 4,

R^(b) is the same or different and each:

(1) C₁₋₆ alkyl, or(2) halogen atom,

n2 is an integer selected from 0 to 4,

m1 is an integer selected from 0 to 3,

m2 is an integer selected from 1 to 4,

X^(a)═X^(b) is:

(1) CH═CH, (2) N═CH, or (3) CH═N,

X is:

(1) nitrogen atom, or(2) C—R^(a) wherein R^(d) is hydrogen atom or halogen atom,

R^(c) is a group selected from the following (1) to (6):

(1) hydrogen atom,(2) C₁₋₆ alkyl optionally substituted by the same or different 1 to 5substituents selected from the following Group A,

(3) —C(═O)—R^(c1), (4) —C(═O)—O—R^(c2),

(5) —C(═O)—NR^(c3)R^(c4) in which R^(c1), R^(c2), R^(c3) and R^(c4) arethe same or different and each:(i) hydrogen atom, or(ii) C₁₋₆ alkyl optionally substituted by the same or different 1 to 5substituents selected from the following Group A, or(6) a group of formula:

in which Y is a group selected from the following (i) to (iii):(i) C₁₋₆ alkylene,

(ii) —C(═O)—, or

(iii) —C(═O)—O—,

Ring T is:

(i) C₆₋₁₀ aryl,(ii) C₃₋₁₀ cycloalkyl, or(iii) saturated monoheterocyclyl wherein the saturated monoheterocyclylcomprises 1 to 4 heteroatoms selected from nitrogen atom, oxygen atom orsulfur atom as well as carbon atoms and the number of the constituentring atoms is 3 to 7,

R^(c5) is the same or different and each:

(i) cyano, or(ii) nitro,

p is an integer selected from 0 to 4,

Group A is the group consisting of:

(a) hydroxyl,(b) C₁₋₆ alkoxy,(c) cyano,(d) C₁₋₆ alkoxycarbonyl,(e) C₁₋₆ alkylcarbonyloxy, and(f) C₂₋₆ alkenyloxy, or a pharmaceutically acceptable salt thereof, or asolvate thereof.[2] The compound of [1], wherein, in the general formula [I], n1 is aninteger selected from 0 to 2,

n2 is an integer selected from 0 to 2,

m1 is an integer selected from 0 to 3,

m2 is an integer selected from 1 to 3,

X is:

(1) nitrogen atom, or(2) C—R^(d) wherein R^(d) is halogen atom,

R^(c) is a group selected from the following (1) to (6):

(1) hydrogen atom,(2) C₁₋₆ alkyl substituted by one substituent selected from thefollowing Group A,

(3) —C(═O)—R^(c1), (4) —C(═O)—O—R^(c2),

(5) —C(═O)—NR^(c3)R^(c4)in which R^(c) is C₁₋₆ alkyl optionally substituted by one substituentselected from the following Group A,

R^(c2) is C₁₋₆ alkyl,

R^(c3) is C₁₋₆ alkyl optionally substituted by one substituent selectedfrom the following Group A,

R^(c4) is

(i) hydrogen atom, or(ii) C₁₋₆ alkyl, or(6) a group of formula:

in which Y^(a) is a group selected from the following (i) to (iii):(i) C₁₋₆ alkylene,

(ii) —C(═O)—, or

(iii) —C(═O)—O—,

Ring T is:

(i) phenyl,(ii) C₃₋₆ cycloalkyl, or(iii) pyrrolidinyl,

R^(c5) is

(i) cyano, or(ii) nitro,

p is an integer selected from 0 or 1,

Group A is the group consisting of:

(a) hydroxyl,(b) C₁₋₆ alkoxy,(c) cyano,(d) C₁₋₆ alkoxycarbonyl,(e) C₁₋₆ alkylcarbonyloxy, and(f) C₂₋₆ alkenyloxy, or a pharmaceutically acceptable salt thereof, or asolvate thereof.[3] The compound of either one of [1] or [2], wherein m1 is an integerof 0 or 1 and m2 is an integer of 1 or 2, or a pharmaceuticallyacceptable salt thereof, or a solvate thereof.[4] The compound of [3], wherein a combination of (m1,m2) is (1,2),which is a compound of the general formula [II]:

wherein each symbol has the same meaning as defined in [1], or apharmaceutically acceptable salt thereof, or a solvate thereof.[5] The compound of [3], wherein a combination of (m1,m2) is (0,2),which is a compound of the general formula [III]:

wherein each symbol has the same meaning as defined in [1], or apharmaceutically acceptable salt thereof, or a solvate thereof.[6] The compound of [3], wherein a combination of (m1,m2) is (0,1),which is a compound of the general formula [IV]:

wherein each symbol has the same meaning as defined in [1], or apharmaceutically acceptable salt thereof, or a solvate thereof.[7] The compound of either one of [1] or [2], wherein a combination of(m1,m2) is selected from (0,3), (2,1), (2,2) or (3,2), or apharmaceutically acceptable salt thereof, or a solvate thereof.[8] The compound of any one of [1] to [7], wherein X^(a)═X^(b) is CH═CHand X is nitrogen atom, or a pharmaceutically acceptable salt thereof,or a solvate thereof.[9] The compound of either one of [1] or [2], wherein a combination of(n1,n2) is (0,0), or a pharmaceutically acceptable salt thereof, or asolvate thereof.[10] The compound of either one of [1] or [2], wherein a combination of(n1,n2) is (1,0), or a pharmaceutically acceptable salt thereof, or asolvate thereof.[11] The compound of either one of [1] or [2], wherein a combination of(n1,n2) is (0,1), or a pharmaceutically acceptable salt thereof, or asolvate thereof.[12] The compound of either one of [1] or [2], wherein a combination of(n1,n2) is (2,0), or a pharmaceutically acceptable salt thereof, or asolvate thereof.[13] The compound of either one of [1] or [2], wherein a combination of(n1,n2) is (0,2), or a pharmaceutically acceptable salt thereof, or asolvate thereof.[14] The compound of either one of [10] or [12], wherein R^(a) is methylor fluorine atom, or a pharmaceutically acceptable salt thereof, or asolvate thereof.[15] The compound of any one of [1] to [14], wherein R^(c) is—C(═O)—R^(c1), or a pharmaceutically acceptable salt thereof, or asolvate thereof.[16] The compound of [15], wherein R^(c) is C₁₋₆ alkyl substituted byone hydroxyl or cyano group, or a pharmaceutically acceptable saltthereof, or a solvate thereof.[17] The compound of any one of [1] to [14], wherein R^(c) is—C(═O)—NR^(c3)R^(c4), or a pharmaceutically acceptable salt thereof, ora solvate thereof.[18] The compound of [17], wherein R^(c3) is C₁₋₆ alkyl substituted byone cyano group, and R^(c4) is hydrogen, or a pharmaceuticallyacceptable salt thereof, or a solvate thereof.[19] The compound of [1] selected from the following chemical structuralformula:

or a pharmaceutically acceptable salt thereof, or a solvate thereof.[20] The compound of [1] selected from the following chemical structuralformula:

or a pharmaceutically acceptable salt thereof, or a solvate thereof.[21] A pharmaceutical composition, comprising the compound of any one of[1] to [20] or a pharmaceutically acceptable salt thereof, or a solvatethereof, and a pharmaceutically acceptable carrier.[22] A Janus kinase inhibitor, comprising the compound of any one of [1]to [20] or a pharmaceutically acceptable salt thereof, or a solvatethereof.[23] The Janus kinase inhibitor of [22], wherein the Janus kinase isJanus kinase 3.[24] The Janus kinase inhibitor of [22], wherein the Janus kinase isJanus kinase 2.[25] A therapeutic or preventive agent for a disease selected from thegroup consisting of organ transplant rejection, graft versus hostreaction after transplantation, autoimmune disease, allergic disease andchronic myeloproliferative disease, comprising the compound of any oneof [1] to [20] or a pharmaceutically acceptable salt thereof, or asolvate thereof.[26] A therapeutic or preventive agent for rheumatoid arthritis,comprising the compound of any one of [1] to [20] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof.[27] A therapeutic or preventive agent for psoriasis, comprising thecompound of any one of [1] to [20] or a pharmaceutically acceptable saltthereof, or a solvate thereof.[28] A method for inhibiting Janus kinase, comprising administering to amammal a pharmaceutically effective amount of the compound of any one of[1] to [20] or a pharmaceutically acceptable salt thereof, or a solvatethereof.[29] The method of [28], wherein the Janus kinase is Janus kinase 3.[30] The method of [28], wherein the Janus kinase is Janus kinase 2.[31] A method for treating or preventing a disease selected from thegroup consisting of organ transplant rejection, graft versus hostreaction after transplantation, autoimmune disease, allergic diseasesand chronic myeloproliferative disease, comprising administering to amammal a pharmaceutically effective amount of the compound of any one of[1] to [20] or a pharmaceutically acceptable salt thereof, or a solvatethereof.[32] A method for treating or preventing rheumatoid arthritis,comprising administering to a mammal a pharmaceutically effective amountof the compound of any one of [1] to [20] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof.[33] A method for treating or preventing psoriasis, comprisingadministering to a mammal a pharmaceutically effective amount of thecompound of any one of [1] to [20] or a pharmaceutically acceptable saltthereof, or a solvate thereof.[34] Use of the compound of any one of [1] to [20] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof in the manufacture of aJanus kinase inhibitor.[35] The use of [34], wherein the Janus kinase is Janus kinase 3.[36] The use of [34], wherein the Janus kinase is Janus kinase 2.[37] Use of the compound of any one of [1] to [20] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof in the manufacture of amedicament for treating or preventing a disease selected from the groupconsisting of organ transplant rejection, graft versus host reactionafter transplantation, autoimmune disease, allergic diseases and chronicmyeloproliferative disease.[38] Use of the compound of any one of [1] to [20] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof in the manufacture of amedicament for treating or preventing rheumatoid arthritis.[39] Use of the compound of any one of [1] to [20] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof in the manufacture of amedicament for treating or preventing psoriasis.[40] A commercial kit, comprising (a) a pharmaceutical compositioncomprising as the active ingredient the compound of any one of [1] to[20] or a pharmaceutically acceptable salt thereof, or a solvatethereof; and (b) a drug package insert of the pharmaceutical compositionwhich indicates that the pharmaceutical composition may be used orshould be used for the treatment or prevention of rheumatoid arthritisor psoriasis.[41] A commercial package, comprising (a) a pharmaceutical compositioncomprising as the active ingredient the compound of any one of [1] to[20] or a pharmaceutically acceptable salt thereof, or a solvatethereof; and (b) a drug package insert of the pharmaceutical compositionwhich indicates that the pharmaceutical composition may be used orshould be used for the treatment or prevention of rheumatoid arthritisor psoriasis.

Effect of Invention

The inventive nitrogen-containing spiro cyclic compound of the presentapplication is effective as a therapeutic or preventive agent for organtransplant rejection, graft versus host reaction after transplantation,autoimmune disease and allergic disease, etc. due to its JAK3activity-inhibition effect.

Moreover, the inventive nitrogen-containing spiro cyclic compound of thepresent application is effective as a therapeutic or preventive agentfor chronic myeloproliferative disease due to its JAK2activity-inhibition effect.

BEST MODE FOR CARRYING OUT THE INVENTION

Terms and phrases used herein are defined as below.

The phrase “optionally substituted” includes both cases thatsubstitutable positions are substituted and are not substituted(unsubstituted) in the subject group. The term “unsubstituted” refers tosuch a case that all substitutable positions are substituted by hydrogenatom in the subject group.

For example, “C₁₋₆ alkyl optionally substituted by the same or different1 to 5 substituents selected from Group A” includes both cases thatsubstitutable positions of C₁₋₆ alkyl are substituted by the same ordifferent 1 to 5 substituents selected from Group A and not substituted(unsubstituted).

The phrase “halogen atom” includes fluorine atom, chlorine atom, bromineatom or iodine atom, preferably fluorine atom or chlorine atom.

The phrase “C₁₋₆ alkyl” refers to C₁₋₆ straight- or branched-chainsaturated hydrocarbon, e.g. methyl, ethyl, propyl, isopropyl, butyl,isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl,1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl,3,3-dimethylbutyl, 2-ethylbutyl, preferably methyl, ethyl, propyl,isopropyl, etc.

The phrase “C₂₋₆ alkenyl” refers to C₂₋₆ straight- or branched-chainunsaturated hydrocarbon containing one or more double bonds, e.g. vinyl,1-methylvinyl, 1-propenyl, allyl, methylpropenyl (including1-methyl-1-propenyl, 2-methyl-1-propenyl, etc.), 1-butenyl, 2-butenyl,3-butenyl, methylbutenyl (including 1-methyl-1-butenyl,2-methyl-1-butenyl, 3-methyl-1-butenyl, etc.), pentenyl, methylpentenyl,hexenyl, preferably vinyl, 1-methylvinyl, 1-propenyl, methylpropenyl,etc.

The phrase “C₁₋₆ alkylene” refers to a divalent group derived fromstraight-chain C₁₋₆ alkyl as defined above, e.g. methylene, ethylene,trimethylene, tetramethylene, pentamethylene, hexamethylene, preferablymethylene, ethylene, etc.

The phrase “C₆₋₁₀ aryl” refers to C₆₋₁₀ aromatic hydrocarbon, e.g.phenyl, 1-naphthyl, 2-naphthyl, preferably phenyl.

The phrase “C₃₋₁₀ cycloalkyl” refers to C₃₋₁₀ monocyclic saturatedhydrocarbon, e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, preferably C₃₋₆ cycloalkyl (includingcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.).

The phrase “saturated monoheterocyclyl” wherein the saturatedmonoheterocyclyl comprises 1 to 4 heteroatoms selected from nitrogenatom, oxygen atom or sulfur atom as well as carbon atoms and the numberof the constituent ring atoms is 3 to 7 includes oxyranyl, thiolanyl,aziridinyl, azetidinyl, oxetanyl, pyrrolidinyl, pyrrolidino (including1-pyrrolidinyl), tetrahydrofuranyl, tetrahydrothienyl, oxazolinyl,oxazolidinyl, isoxazolinyl, isoxazolidinyl, thiazolinyl, thiazolidinyl,isothiazolinyl, isothiazolidinyl, imidazolinyl, imidazolidinyl,pyrazolinyl, pyrazolidinyl, piperidinyl, piperidino (including1-piperidinyl), morpholinyl, morpholino (including 4-morpholinyl),thiomorpholinyl, thiomorpholino (including 4-thiomorpholinyl),piperazinyl, piperazino (including 1-piperazinyl),hexahydro-1,3-oxazinyl, homomorpholine, homopiperazine, etc.

The phrase “C₁₋₆ alkoxy” refers to C₁₋₆ straight- or branched-chainalkoxy, particularly methoxy, ethoxy, propoxy, isopropoxy, butoxy,isobutoxy, s-butoxy, t-butoxy, pentyloxy, isopentyloxy, 2-methylbutoxy,neopentyloxy, 1-ethylpropoxy, hexyloxy, etc.

The phrase “C₁₋₆ alkoxycarbonyl” refers to a group wherein C₁₋₆straight- or branched-chain alkoxy binds to carbonyl, particularlymethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl,butoxycarbonyl, isobutoxycarbonyl, s-butoxycarbonyl, t-butoxycarbonyl,pentyloxycarbonyl, isopentyloxycarbonyl, 2-methylbutoxycarbonyl,neopentyloxycarbonyl, 1-ethylpropoxycarbonyl, hexyloxycarbonyl,4-methylpentyloxycarbonyl, etc.

The phrase “C₁₋₆ alkylcarbonyloxy” refers to a group wherein a “groupwherein C₁₋₆ alkyl binds to carbonyl” binds to oxy, particularlyacetyloxy, propionyloxy, butyryloxy, isobutyryloxy, etc.

The phrase “C₂₋₆ alkenyloxy” refers to a group wherein “C₂₋₆ alkenyl”binds to oxy, particularly allyloxy, 1-butenyloxy, etc.

Preferable embodiments for each group in a compound of the generalformula [I], which is also referred to as the present compoundhereinafter, are illustrated as below.

Preferable embodiments of R^(a) include:

(1) methyl, or(2) fluorine atom, etc.

Preferable embodiments of n1 include an integer of 0, 1 or 2.

Preferable embodiments of R^(b) include:

(1) methyl, or(2) fluorine atom, etc.

Preferable embodiments of n2 include an integer of 0, 1 or 2.

Preferable embodiments of m1 include an integer selected from 0 to 3.

Preferable embodiments of m2 include an integer of 1, 2 or 3.

A combination of (m1,m2) includes (0,1), (0,2), (0,3), (1,1), (1,2),(2,1), (2,2) or (3,2).

Preferably, R^(a) and R^(b) may be substituted on carbon atomsconstituting each spirocycle of the general formula [I] except for aspiro carbon, and carbon atoms which are not substituted by R^(a) orR^(b) are saturated by hydrogen atoms. In case that n1 is 2 or above,R^(a) may be the same or different and each, and substituted on the sameor different positions. Further, in case that n2 is 2 or above, R^(b)may be the same or different and each, and substituted on the same ordifferent positions.

X^(a)═X^(b) is:

(1) CH═CH, (2) N═CH, or (3) CH═N,

preferably (1) CH═CH.

Preferable embodiments of X are:

(1) nitrogen atom, or

(2) C—Cl,

more preferably (1) nitrogen atom.

Preferable embodiments of R^(c) include hydrogen atom, cyanoethyl,acetyl, benzyl, cyanomethylcarbonyl, propenyloxyethylcarbonyl,2-propanylcarbonyl, ethylcarbonyl, methoxycarbonyl,(S)-hydroxyethylcarbonyl, hydroxymethylcarbonyl, 1-hydroxyethylcarbonyl,acetoxymethylcarbonyl, (S)-acetoxyethylcarbonyl, methoxymethylcarbonyl,methoxyethylcarbonyl, (S)-methoxyethylcarbonyl,(R)-methoxyethylcarbonyl, 3-cyanopyrrolidinylcarbonyl,3-cyanophenylcarbonyl, 4-cyanophenylcarbonyl,methoxycarbonylethylcarbonyl, p-nitrophenoxycarbonyl,1-cyanomethylcyclopropanylcarbonyl, t-butoxycarbonyl, N-ethylcarbamoyl,N-cyanomethylcarbamoyl, N-cyanoethylcarbamoyl,N,N-methylcyanomethylcarbamoyl, N,N-methylcyanoethylcarbamoyl,N-propanylcarbamoyl, preferably cyanomethylcarbonyl,hydroxymethylcarbonyl or cyanoethylcarbamoyl.

Preferable embodiments of a compound of the general formula [I] includecompounds of the following formulae:

A compound of the general formula [II] includes preferably a compound ofthe general formula [II-A], [II-B] or [II-C].

A compound of the general formula [III] includes preferably a compoundof the general formula [III-A], [III-B] or [III-C].

A compound of the general formula [IV] includes preferably a compound ofthe general formula [IV-A], [IV-B] or [IV-C].

Another preferable embodiment of a compound of the general formula [I]includes compounds of the following formula:

Another preferable embodiment of a compound of the general formula [I]has JAK1 and/or JAK2 and/or Tyk2 inhibitory effects as well as JAK3inhibitory effect. More preferably, a compound of the general formula[I] has all of JAK1, JAK2, JAK3 and Tyk2 inhibitory effects.

The “pharmaceutically acceptable salt” of a compound of the generalformula [I], which is also referred to as the present compoundhereinafter, may be any a toxic salts of the present compound andincludes a salt with an inorganic acid, an organic acid, an inorganicbase, an organic base, an amino acid, etc.

The salt with an inorganic acid includes a salt with hydrochloric acid,nitric acid, sulfuric acid, phosphoric acid, hydrobromic acid, etc.

The salt with an organic acid includes a salt with oxalic acid, maleicacid, citric acid, fumaric acid, lactic acid, malic acid, succinic acid,tartaric acid, acetic acid, trifluoroacetic acid, gluconic acid,ascorbic acid, methanesulfonic acid, benzenesulfonic acid,p-toluenesulfonic acid, etc.

The salt with an inorganic base includes sodium salt, potassium salt,calcium salt, magnesium salt, ammonium salt, etc.

The salt with an organic base includes a salt with methylamine,diethylamine, trimethylamine, triethylamine, ethanolamine,diethanolamine, triethanolamine, ethylenediamine,tris(hydroxymethyl)methylamine, dicyclohexylamine,N,N′-dibenzylethylenediamine, guanidine, pyridine, picoline, choline,cinchonine, meglumine, etc.

The salt with an amino acid includes a salt with lysine, arginine,aspartic acid, glutamic acid, etc.

According to known methods, each salt may be obtained by reacting acompound of the general formula [I] with an inorganic base, an organicbase, an inorganic acid, an organic acid or an amino acid.

The “solvate” refers to a material wherein solvent molecules coordinatewith a compound of the general formula [I] or a pharmaceuticallyacceptable salt thereof, and includes hydrate. A preferable solvate is apharmaceutically acceptable solvate including 1 hydrate, 1/2 hydrate or2 hydrate of a compound of the general formula [I], 1 hydrate of sodiumsalt of a compound of the general formula [I], 1 methanolate, 1ethanolate or 1 acetonitrilate of a compound of the general formula [I],2/3 ethanolate of 2 hydrochloride of a compound of the general formula[I], etc. More preferable solvate is 1 hydrate of a compound of thegeneral formula [I]. According to known methods, a solvate thereof maybe obtained.

A compound of the general formula [I] also exists as various “isomers”.For example, a geometric isomer thereof includes E- and Z-isomers. Incase that any asymmetric carbon atoms exist, a stereoisomer based onsuch carbon atoms includes enantiomers and diastereomers. In case thatany chiral axes exist, a stereoisomer based on such axes exists. Atautomer may exist as the case may be. Therefore, the scope of thepresent invention encompasses all these isomers and a mixture thereof.

A compound of the general formula [I] may be also labelled by isotopes(e.g., ³H, ¹⁴C, ³⁵S, etc.).

A preferable compound of the general formula [I] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof is substantially-purifiedcompound of the general formula [I] or a pharmaceutically acceptablesalt thereof, or a solvate thereof. More preferable one is a compound ofthe general formula [I] or a pharmaceutically acceptable salt thereof,or a solvate thereof which is purified in the purity of 80% or above.

In the present invention, a prodrug of a compound of the general formula[I] may be also a useful drug. The “prodrug” refers to a derivative ofthe present compound with chemically or metabolically decomposablefunctional groups which is administered in vivo, followed by convertinginto a corresponding parent compound by hydrolysis, solvolysis orphysiological decomposition to show original drug efficacies, andincludes any composites with noncovalent bonds and salts thereof. Theprodrug is utilized to improve absorption in oral administration or totarget at its target site, for example. Modification sites for theprodrug formation include any reactive functional groups of the presentcompound including hydroxyl, carboxyl, amino, thiol, etc.

A modified group for hydroxyl includes acetyl, propionyl, isobutyryl,pivaloyl, palmitoyl, benzoyl, 4-methylbenzoyl, dimethylcarbamoyl,dimethylaminomethylcarbonyl, sulfo, alanyl, fumaryl, or sodium-saltified3-carboxybenzoyl or 2-carboxyethylcarbonyl, etc.

A modified group for carboxyl includes methyl, ethyl, propyl, isopropyl,butyl, isobutyl, tert-butyl, pivaloyloxymethyl, carboxymethyl,dimethylaminomethyl, 1-(acetyloxy)ethyl, 1-(ethoxycarbonyloxy)ethyl,1-(isopropyloxycarbonyloxy)ethyl, 1-(cyclohexyloxycarbonyloxy)ethyl,(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl, benzyl, phenyl, o-tolyl,morpholinoethyl, N,N-diethylcarbamoylmethyl, phthalidyl, etc.

A modified group for amino includes tert-butyl, docosanoyl,pivaloylmethyloxy, alanyl, hexylcarbamoyl, pentylcarbamoyl,3-methylthio-1-(acetylamino)propylcarbonyl,1-sulfo-1-(3-ethoxy-4-hydroxyphenyl)methyl,(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl,(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl, tetrahydrofuranyl,pyrrolidylmethyl, etc.

The “pharmaceutical composition” includes an oral preparation such astablet, capsule, granule, powder, lozenge, syrup, emulsion, suspension,or a parenteral preparation such as external preparation, suppository,injection, drop, nasal drug, pulmonary drug.

The pharmaceutical composition of the present invention may be preparedby properly mixing a compound of the general formula [I] or apharmaceutically acceptable salt thereof, or a solvate thereof with atleast one or more types of pharmaceutically acceptable carriers inappropriate amounts according to known methods in the medicinalpreparation field. A content of a compound of the general formula [I] ora pharmaceutically acceptable salt thereof, or a solvate thereof in thepharmaceutical composition depends on its dosage forms, dosage amounts,etc., and for example, is 0.1 to 100% by weight of the composition.

The “pharmaceutically acceptable carrier” includes various conventionalorganic or inorganic carriers for pharmaceutical materials, e.g.,excipient, disintegrant, binder, fluidizer, lubricant for solidpreparations, or solvent medium, solubilizing agent, suspending agent,tonicity agent, buffer, soothing agent for liquid preparations. Further,an additive including a preserving agent, an antioxidant agent, acolorant, a sweetening agent may be used, if needed.

The “excipient” includes lactose, sucrose, D-mannitol, D-sorbitol,cornstarch, dextrin, microcrystalline cellulose, crystalline cellulose,carmellose, carmellose calcium, sodium carboxymethyl starch,low-substituted hydroxypropylcellulose, gum arabic, etc.

The “disintegrant” includes carmellose, carmellose calcium, carmellosesodium, sodium carboxymethyl starch, croscarmellose sodium,crospovidone, low-substituted hydroxypropylcellulose,hydroxypropylmethylcellulose, crystalline cellulose, etc.

The “binder” includes hydroxypropylcellulose,hydroxypropylmethylcellulose, povidone, crystalline cellulose, sucrose,dextrin, starch, gelatin, carmellose sodium, gum arabic, etc.

The “fluidizer” includes light anhydrous silicic acid, magnesiumstearate, etc.

The “lubricant” includes magnesium stearate, calcium stearate, talc,etc.

The “solvent medium” includes purified water, ethanol, propylene glycol,macrogol, sesame oil, corn oil, olive oil, etc.

The “solubilizing agent” includes propylene glycol, D-mannitol, benzylbenzoate, ethanol, triethanolamine, sodium carbonate, sodium citrate,etc.

The “suspending agent” includes benzalkonium chloride, carmellose,hydroxypropylcellulose, propylene glycol, povidone, methylcellulose,glyceryl monostearate, etc.

The “tonicity agent” includes glucose, D-sorbitol, sodium chloride,D-mannitol, etc.

The “buffer” includes sodium hydrogen phosphate, sodium acetate, sodiumcarbonate, sodium citrate, etc.

The “soothing agent” includes benzyl alcohol, etc.

The “preserving agent” includes ethyl paraoxybenzoate, chlorobutanol,benzyl alcohol, sodium dehydroacetate, sorbic acid, etc.

The “antioxidant agent” includes sodium sulfite, ascorbic acid, etc.

The “colorant” includes food dye (e.g., Food Red No. 2 or 3, Food YellowNo. 4 or 5, etc.), 3-carotene, etc.

The “sweetening agent” includes saccharin sodium, dipotassiumglycyrrhizinate, aspartame, etc.

The pharmaceutical composition of the present invention may be orally orparenterally (e.g., locally, rectally, intravenously, etc.) administeredto a mammal except human beings (e.g., mice, rats, hamsters, guineapigs, rabbits, cats, dogs, pigs, cows, horses, sheep, monkeys, etc.) aswell as human beings. A dose of said pharmaceutical composition dependson administration subjects, diseases, conditions, dosage forms,administration routes, and the dose in case of orally administering toadult patients (body weight: about 60 kg) who are suffering from organtransplant rejection, graft versus host reaction after transplantation,autoimmune disease or allergic disease, etc. is usually in the rangefrom about 1 mg to 1 g per day with comprising the present compound asthe active ingredient. The dose may be administered at a time or inseveral divided doses.

The compound of the general formula [I] or a pharmaceutically acceptablesalt thereof, or a solvate thereof may be used as the active ingredientof a therapeutic or preventive agent for the following diseases due toits JAK3 inhibitory activity:

(a) organ transplant rejection, or graft versus host reaction aftertransplantation;(b) autoimmune diseases including rheumatoid arthritis, psoriasis,psoriatic arthritis, multiple sclerosis, ulcerative colitis, Crohn'sdisease, systemic lupus erythematosus, type I diabetes, myastheniagravis, Castleman's disease, juvenile idiopathic arthritis, dry eye; and(c) allergic diseases including asthma, atopic dermatitis, rhinitis.

Preferably, a compound of the general formula [I] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof may be used as the activeingredient of a therapeutic or preventive agent for rheumatoid arthritisor psoriasis.

Moreover, a compound of the general formula [I] or a pharmaceuticallyacceptable salt thereof, or a solvate thereof may be used as the activeingredient of a therapeutic or preventive agent for chronicmyeloproliferative diseases including polycythemia vera, primarymyelofibrosis, essential thrombocythemia, etc. due to its JAK2inhibitory activity.

The phrase “JAK inhibitory” refers to inhibiting functions of JAK todisappear or reduce its activity, and inhibiting any functions of JAKfamily. A preferable “JAK inhibitory” is “human JAK inhibitory”.

A preferable “JAK inhibitor” is “human JAK inhibitor”.

The phrase “JAK3 inhibitory” refers to inhibiting functions of JAK3 todisappear or reduce its activity. For example, it refers to inhibitingfunctions of JAK3 under the conditions of test examples as describedhereinbelow. A preferable “JAK3 inhibitory” is “human JAK3 inhibitory”.

A preferable “JAK3 inhibitor” is “human JAK3 inhibitor”.

The phrase “JAK2 inhibitory” refers to inhibiting functions of JAK2 todisappear or reduce its activity. For example, it refers to inhibitingfunctions of JAK2 under the conditions of test examples as describedhereinbelow. A preferable “JAK2 inhibitory” is “human JAK2 inhibitory”.

A preferable “JAK2 inhibitor” is “human JAK2 inhibitor”.

As an example, a method for preparing compounds for working the presentinvention is illustrated as follows, and the method for preparing thepresent compound is not intended to be limited thereto.

Unless otherwise specified, effective preparation methods may be carriedout by introducing any protecting groups to any functional groups, ifneeded, and then deprotecting such groups at a later step; treating anyfunctional groups in the forms of their precursors in each step andconverting such precursors into the corresponding desirable functionalgroups at an appropriate step; or interchanging each order of processand step.

In each step, each aftertreatment of reaction may be carried out in aconventional manner, and an isolation and purification process may beoptionally selected from the conventional method includingcrystallization, recrystallization, distillation, separation, silica gelchromatography, preparative HPLC, or a combination thereof. The roomtemperature refers to 1° C. to 40° C.

General Preparation Method 1 General Preparation Method of Compound [I]

[In the above scheme, P^(N) is a protecting group of amine, preferablytert-butoxycarbonyl, benzyl, p-methoxybenzyl, benzyloxycarbonyl; Q is N(nitrogen atom) substituted by a protecting group, or NH; Hal is ahalogen atom; R^(a), R^(b), n1, n2, X^(a), X^(b), X, m1, m2 and R^(c)have the same meanings as defined in the above formula [I].]

(Step 1)

Compound [Va] may be reacted with Compound [Vb] in a solvent in thepresence of a base to give Compound [Vc].

The solvent used in the reaction includes an ester solvent such as ethylacetate; a ketone solvent such as acetone; an amide solvent such asN,N-dimethylformamide; an alcohol solvent such as ethanol; an ethersolvent such as dioxane; a hydrocarbon solvent such as toluene; ahalogenated hydrocarbon solvent such as chloroform; water, and may beused alone or in a combination of 2 or more of them. A preferablesolvent in the reaction is water.

The base used in the reaction includes triethylamine,N,N-diisopropylethylamine, pyridine, 4-dimethylaminopyridine,N-methylmorpholine, lithium hydroxide, sodium hydroxide, potassiumhydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate,preferably potassium carbonate.

The reaction temperature is usually room temperature to 110° C.,preferably about 80° C. to 110° C.

The reaction time is usually about 30 minutes to 3 days, preferablyabout 3 hours to 1 day.

(Step 2)

Compound [Vd] may be obtained by removing P^(N) of Compound [Vc] in aconventional manner of amine-deprotection reaction. The deprotectionreaction may be carried out by known methods depending on selectedprotecting groups.

For example, Compound [Vc] wherein P^(N) is tert-butoxycarbonyl may betreated in a single or mixed solvent of chloroform, dioxane, ethylacetate, ethanol, methanol, water, etc. with an acid includinghydrochloric acid, trifluoroacetic acid.

For example, Compound [Vc] wherein P^(N) is benzyl or benzyloxycarbonylmay be hydrogenated in a single or mixed solvent of chloroform,tetrahydrofuran, dioxane, ethyl acetate, ethanol, methanol, etc. in thepresence of a catalyst including palladium carbon, palladium hydroxide.

(Step 3)

Compound [Vd] may be introduced R^(c) in a solvent to give Compound [I].

For example, Compound [Vd] may be reacted with1-cyanoacetyl-3,5-dimethylpyrazole in a solvent in the presence of abase to give Compound [I] wherein R^(c) is cyanoacetyl. The solvent usedin the reaction includes an ester solvent such as ethyl acetate; aketone solvent such as acetone; an amide solvent such asN,N-dimethylformamide; an alcohol solvent such as ethanol; an ethersolvent such as dioxane; a hydrocarbon solvent such as toluene; ahalogenated hydrocarbon solvent such as chloroform, and may be usedalone or in a combination of 2 or more of them. A preferable solvent inthe reaction is dioxane. The base used in the reaction includestriethylamine, pyridine, 4-dimethylaminopyridine, N-methylmorpholine,N,N-diisopropylethylamine, preferably N,N-diisopropylethylamine. Thereaction temperature is usually room temperature to 110° C., preferablyabout 80° C. to 110° C. The reaction time is usually about 30 minutes to1 day, preferably about 2 to 4 hours.

For example, Compound [Vd] may be reacted with a carboxylic acidcompound in a conventional amidation reaction in a solvent in thepresence of a condensing agent and a base to give Compound [I] whereinR^(c) is acyl. The solvent used in the reaction includes an amidesolvent such as N,N-dimethylformamide; a halogenated hydrocarbon solventsuch as chloroform, and may be used alone or in a combination of 2 ormore of them. A preferable solvent in the reaction is an ether solventsuch as tetrahydrofuran or dioxane. The condensing agent used in thereaction includes water-soluble carbodiimide (e.g.,1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide hydrochloride),N,N′-dicyclohexylcarbodiimide, diphenylphosphoryl azide,carbonyldiimidazole. To the reaction mixture may be added1-hydroxy-1H-benzotriazole, 4-dimethylaminopyridine, if needed. Apreferable condensing agent in the reaction is carbonyldiimidazole.

In the above amidation reaction, the carboxylic acid compound may bealso pre-converted into a corresponding acid chloride or mixed acidanhydride, and then reacted with Compound [Vd] to give Compound [I].

For example, Compound [Vd] may be reacted with acrylonitrile in thepresence of a base including triethylamine, pyridine,N,N-diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene in anamide solvent including N,N-dimethylformamide, N,N-dimethylacetamide, oracetonitrile to give Compound [I] wherein R^(c) is propylnitrile.

For example, Compound [Vd] may be reacted with alkyl chloroformate, etc.by a conventional synthesis of carbamate to give Compound [I] whereinR^(c) is alkoxycarbonyl.

For example, Compound [Vd] may be reacted with alkyl isocyanate, etc. bya conventional synthesis of urea to give Compound [I] wherein R^(c) isalkylaminocarbonyl.

In the above urea synthesis, the alkylamine compound may be also reactedwith 4-nitrophenyl chloroformate to give alkyl carbamic acid4-nitrophenyl ester which may be sequentially reacted with Compound [Vd]to give Compound [I].

A conventional deprotection of amine in Compound [Vd] wherein N of Q issubstituted by a protecting group may be optionally carried out after orbefore introducing R^(c). The deprotection may be carried out by knownmethods depending on selected protecting groups.

For example, the deprotection in which a protecting group isp-toluenesulfonyl may be carried out by treating with an alkaliincluding sodium hydroxide, potassium hydroxide, cesium carbonate in asingle or mixed solvent of an ether solvent such as tetrahydrofuran ordioxane; an alcohol solvent such as ethanol or methanol; water, etc.

For example, the deprotection in which a protecting group isp-methoxybenzyl may be carried out by treating with an acid such ashydrochloric acid, trifluoroacetic acid in a single or mixed solvent ofan ether solvent such as anisole; a halogenated hydrocarbon solvent suchas chloroform; an ester solvent such as ethyl acetate; an ether solventsuch as dioxane; an alcohol solvent such as ethanol or methanol; water,etc.

As an example, some synthetic methods of Compound [Va] in GeneralPreparation Method 1 are illustrated in the following GeneralPreparation Methods 2 to 4.

In the following General Preparation Methods 2 to 4, Compounds [VIi] and[VIIo] correspond to Compound [Va].

General Preparation Method 2

[In the above scheme, P^(N1), P^(N2) and P^(N3) are a protecting groupof amine, preferably tert-butoxycarbonyl, benzyl, benzyloxycarbonyl; Yis hydroxyl, or a leaving group including chlorine atom, bromine atom,iodine atom, mesyloxy, tosyloxy; m1′ is 0 or 1; carbon atoms with * maybe substituted by R^(a) so as to be chemically acceptable; R^(a), R^(b),n1, n2 and m2 have the same meanings as defined in the above formula[I].]

(Step 1a)

Compound [VIa] may be conventionally esterified with Compound [VIb] in asolvent to give Compound [VIc]. For example, Compound [VIa] may bereacted with Compound [VIb] wherein Y is hydroxyl in a solvent in thepresence of a condensing agent and a base.

The solvent used in the reaction includes an amide solvent such asN,N-dimethylformamide; an ether solvent such as tetrahydrofuran; ahalogenated hydrocarbon solvent such as chloroform, and may be usedalone or in a combination of 2 or more of them. A preferable solvent inthe reaction is a halogenated hydrocarbon solvent such as chloroform.

A preferable condensing agent used in the reaction is water-solublecarbodiimide (e.g., 1-ethyl-3-(3′-dimethylaminopropyl)carbodiimidehydrochloride), etc.

A preferable base used in the reaction is an organic base such as4-dimethylaminopyridine, etc.

A preferable reaction temperature is room temperature.

The reaction time is usually about 30 minutes to 1 day, preferably about2 to 6 hours.

For example, Compound [VIa] may be also reacted with Compound [VIb]wherein Y is a leaving group in a solvent in the presence of a base.

The leaving group includes chlorine atom, bromine atom, iodine atom,mesyloxy and tosyloxy, preferably bromine atom.

The solvent used in the reaction includes an amide solvent such asN,N-dimethylformamide; an ether solvent such as dioxane; a hydrocarbonsolvent such as toluene, and may be used alone or in a combination of 2or more of them. A preferable solvent in the reaction isN,N-dimethylformamide.

The base used in the reaction includes an inorganic base such as sodiumcarbonate, potassium carbonate, sodium phosphate, potassium phosphate,sodium bicarbonate, preferably potassium carbonate.

The reaction temperature is usually room temperature to 120° C.,preferably room temperature to 60° C.

The reaction time is usually about 30 minutes to 1 day, preferably about2 to 6 hours.

(Step 2a)

Compound [VIc] may be treated by a conventional Claisen rearrangement ina solvent in the presence of a base to give Compound [VId].

A preferable solvent used in the reaction is tetrahydrofuran.

The base used in the reaction includes a base such as lithiumdiisopropylamide, lithium hexamethyl disilazide, potassium hexamethyldisilazide, sodium hexamethyl disilazide, sodium hydride, potassiumtert-butoxide, preferably lithium hexamethyl disilazide.

A preferable reaction temperature is about −80° C. to 0° C.

The reaction time is usually about 30 minutes to 1 day, preferably about2 to 4 hours.

(Step 3a)

The carboxylic acid moiety of Compound [VId] may be converted into acarbamate in a solvent to give Compound [VIe]. The conversion includes aconventional Curtius rearrangement.

A preferable reagent used in the reaction is diphenylphosphoryl azide.

The solvent used in the reaction includes an amide solvent such asN,N-dimethylformamide; an alcohol solvent such as benzyl alcohol; anether solvent such as dioxane; a hydrocarbon solvent such as toluene,and may be used alone or in a combination of 2 or more of them. Apreferable solvent in the reaction is a mixed solution of toluene andbenzyl alcohol.

A preferable base used in the reaction is triethylamine.

The reaction temperature is usually room temperature to 110° C.,preferably about 80° C. to 110° C.

The reaction time is usually about 30 minutes to 2 days, preferablyabout 2 hours to 1 day.

An additive including 4-dimethylaminopyridine may be used, if needed.

(Step 4a)

The olefin moiety of Compound [VIe] may be converted into hydroxyl togive Compound [VIf]. The conversion is illustrated in the following Step4a-1 or 2, as an example.

(Step 4a-1)

Compound [VIe] may be treated in a solvent by an ozone oxidation,followed by a reduction to give Compound [VIf]. The ozone oxidation maybe carried out according to a conventional method.

A preferable solvent used in the ozone oxidation is a mixed solution ofchloroform and methanol.

The reaction temperature in the ozone oxidation is usually about −100°C. to 0° C., preferably about −80° C. to −60° C.

The reaction time in the ozone oxidation is usually about 5 minutes to 6hours, preferably about 15 minutes to 3 hours.

A preferable reagent used in the reduction is sodium borohydride.

The reaction temperature in the reduction is usually about −100° C. toroom temperature, preferably about −20° C. to 0° C.

The reaction time in the reduction is usually about 30 minutes to 6hours, preferably about 1 to 3 hours.

(Step 4a-2)

Compound [VIe] may be treated in a solvent by a hydroboration, followedby an oxidation to give Compound [VIf].

The reagent used in the hydroboration includes borane-pyridine complex,borane-dimethyl sulfide complex, 9-borabicyclo[3.3.1]nonane, or asolution of borane-tetrahydrofuran complex in tetrahydrofuran,preferably a solution of borane-tetrahydrofuran complex intetrahydrofuran.

A preferable solvent used in the hydroboration is tetrahydrofuran.

The reaction temperature in the hydroboration is usually about −20° C.to room temperature, preferably 0° C.

A preferable reaction time in the hydroboration is about 1 to 4 hours.

The reagent used in the oxidation includes hydrogen peroxide or sodiumperoxoborate monohydrate, preferably sodium peroxoborate monohydrate.

The reaction temperature in the oxidation is usually about 0° C. to roomtemperature, preferably room temperature.

A preferable reaction time in the oxidation is about 1 hour to 1 day.

(Step 5a)

P^(N2) among P^(N1) and P^(N2) may be selectively removed from Compound[VIf] by a conventional amine deprotection in the similar manner to theabove Step 2 of General Preparation Method 1 to give Compound [VIg]. Thedeprotection may be carried out by known methods depending on selectedprotecting groups.

(Step 6a)

Compound [VIg] may be treated in a solvent by a cyclization, followed byan introduction of P^(N3) to give Compound [VIh]. The cyclization may becarried out by introducing a leaving group into hydroxyl of Compound[VIg] in a solvent in the presence of a base. As an example, Step 6a-1or 2 is illustrated as below.

(Step 6a-1)

Compound [VIg] may be reacted with carbon tetrabromide andtriphenylphosphine (or alternatively methanesulfonyl chloride in placeof the two reagents) in a solvent in the presence of a base to achievean introduction of a leaving group and a cyclization in one step.

A preferable solvent used in the reaction is dichloromethane.

A preferable base used in the reaction is triethylamine.

A preferable reaction temperature is 0° C. to room temperature.

The reaction time is usually about 10 minutes to 24 hours, preferablyabout 30 minutes to 12 hours.

(Step 6a-2)

The introduction of a leaving group and the cyclization may be dividedin two steps.

A preferable reagent used in the introduction of a leaving group ismethanesulfonyl chloride.

A preferable solvent used in the introduction of a leaving group ischloroform.

A preferable base used in the introduction of a leaving group istriethylamine.

The reaction temperature of the introduction of a leaving group isusually about 0° C. to room temperature, preferably 0° C.

A preferable reaction time of the introduction of a leaving group isabout 30 minutes to 2 hours.

A preferable solvent used in the cyclization is N,N-dimethylformamide.

A preferable base used in the cyclization is sodium hydride.

The reaction temperature of the cyclization is usually about 0° C. toroom temperature, preferably 0° C.

A preferable reaction time of the cyclization is about 10 minutes to 2hours.

The cyclized compound may be introduced P^(N3) by a conventional amineprotection to give Compound [VIh]. The amine protection may be carriedout by known methods depending on selected protecting groups.

For example, a compound wherein P^(N3) is benzyloxycarbonyl may beobtained by treating with benzyl chloroformate in a halogenatedhydrocarbon solvent such as chloroform, dichloromethane in the presenceof an organic base such as triethylamine.

Step 5a may be abbreviated, and then, Step 6a-2 may be carried out.

(Step 7a)

P^(N1) among P^(N1) and P^(N3) may be selectively removed from Compound[VIh] by a conventional amine deprotection in the similar manner to theabove Step 2 of General Preparation Method 1 to give Compound [VIi]. Thedeprotection may be carried out by known methods depending on selectedprotecting groups.

Either Compound [Va], [Vc], [Vd] or [I] in General Preparation Method 1or Compounds [VId] to [VIi] in General Preparation Method 2 may beoptically resolved to give an optically-active compound.

The optical resolution includes a method wherein racemic Compound [VId]and an optically-active amine compound are mixed in a solvent, followedby crystallizing as a single diastereomeric salt. The resultingdiastereomeric salt may be desalted in a conventional manner to giveoptically-active Compound [VId]. (+)- or (−)-Isomer of Compound [VId]may be prepared by adopting an appropriate optically-active aminecompound.

The optically-active amine compound includes(S)-(−)-2-amino-3-phenylpropan-1-ol,(R)-(+)-2-amino-3-phenylpropan-1-ol, (S)-(−)-1-(1-naphthyl)ethylamine,(R)-(+)-1-(1-naphthyl)ethylamine, (S)-(+)-2-amino-2-phenyl-ethanol,(R)-(−)-2-amino-2-phenyl-ethanol.

The solvent includes a ketone solvent such as acetone, methyl ethylketone, methyl isobutyl ketone; an ester solvent such as methyl acetate,ethyl acetate, isopropyl acetate, isobutyl acetate; an ether solventsuch as isopropyl ether, 1,2-dimethoxyethane; an alcohol solvent such asmethanol, ethanol, isopropanol; water, and may be used alone or in acombination of 2 or more of them. A preferable solvent includesisopropyl acetate, isopropanol or 1,2-dimethoxyethane.

Additionally, a conventional method for enhancing the optical purity maybe optionally carried out. For example, a recrystallization may berepeated.

An alternative method for the optical resolution includes a methodwherein racemic Compound [I] is treated by a chiral stationary-phasecolumn to separate desirable optically-active Compound [I] from anotherisomer thereof.

General Preparation Method 3

[In the above scheme, P^(N4), P^(N5), P^(N6) and P^(N7) are a protectinggroup of amine, preferably benzyl, tert-butoxycarbonyl orbenzyloxycarbonyl; P^(C1) is a protecting group of carboxylic acid,preferably tert-butyl ester, methyl ester or ethyl ester; Hal is ahalogen atom; m1′ is 0 or 1; carbon atoms with * may be substituted byR^(a) so as to be chemically acceptable; carbon atoms with # may besubstituted by R^(b) so as to be chemically acceptable; R^(a), R^(b),n1, n2 and m2 have the same meanings as defined in the above formula[I].]

(Step 1b)

Compound [VIIa] may be introduced P^(N4) by a conventional amineprotection to give Compound [VIIb]. The amine protection may be carriedout by known methods depending on selected protecting groups.

For example, in case that P^(N4) is benzyl, the protection may becarried out by hydrogenating with benzaldehyde in an alcohol solventsuch as methanol, ethanol in the presence of a palladium catalyst suchas palladium carbon.

(Step 2b)

Compound [VIIb] may be reacted with Compound [VIIc] in a solvent in thepresence of a base to give Compound [VIId].

A preferable solvent used in the reaction is N,N-dimethylformamide.

A preferable base used in the reaction is potassium carbonate.

The reaction temperature is usually room temperature to 120° C.,preferably room temperature to 60° C.

The reaction time is usually about 30 minutes to 2 days, preferablyabout 6 hours to 1 day.

(Step 3b)

Compound [VIId] may be halogenated in a solvent in a conventionalmanner, followed by a rearrangement to give Compound [VIIe].

A preferable reagent used in the halogenation is thionyl chloride.

A preferable solvent used in the halogenation is chloroform.

A preferable reaction temperature of the halogenation is roomtemperature to 60° C.

The reaction time of the halogenation is usually about 30 minutes to 1day, preferably about 1 to 6 hours.

The rearrangement is illustrated in the following Step 3b2.Specifically, Compound [VIIe2] obtained in the halogenation may berearranged in a solvent to give Compound [VIIe3].

A preferable solvent used in the rearrangement is N,N-dimethylformamide.

A preferable reaction temperature of the rearrangement is about 60° C.to 100° C.

A preferable reaction time of the rearrangement is about 30 minutes to 3days.

(Step 4b)

Compound [VIIe] may be intramolecularly cyclized in a solvent in thepresence of a base to give Compound [VIIf].

A preferable base used in the reaction is lithium hexamethyl disilazide.

A preferable solvent used in the reaction is a mixed solution oftetrahydrofuran and hexamethyl phosphoramide.

The reaction temperature is usually about −100° C. to 0° C., preferablyabout −80° C. to 0° C.

The reaction time is usually about 30 minutes to 6 hours, preferablyabout 1 to 2 hours.

(Step 5b)

P^(N4) of Compound [VIIf] may be converted into P^(N5) to give Compound[VIIg].

The removal of P^(N4) may be carried out by a conventional aminedeprotection in the similar manner to the above General PreparationMethod 1 Step 2. The deprotection may be carried out by known methodsdepending on selected protecting groups.

The introduction of P^(N5) may be carried out by a conventional amineprotection in the similar manner to the above General Preparation Method3 Step 1b. The amine protection may be carried out by known methodsdepending on selected protecting groups.

For example, in case that P^(N4) of Compound [VIIf] is benzyl and P^(N5)of Compound [VIIg] is tert-butoxycarbonyl, P^(N4) may be converted intoP^(N5) in one step by hydrogenating Compound [VIIf] with di-tert-butyldicarbonate in a mixed solvent of tetrahydrofuran and methanol in thepresence of a catalyst such as palladium carbon or palladium hydroxide.

(Step 6b)

Compound [VIIg] may be reacted with Compound [VIIh] in a solvent in thepresence of a base to give Compound [VIIi].

A preferable base used in the reaction is lithium hexamethyl disilazide.

A preferable solvent used in the reaction is tetrahydrofuran.

The reaction temperature is usually about −80° C. to room temperature,preferably about −80° C. to 0° C.

The reaction time is usually about 30 minutes to 3 hours, preferablyabout 30 minutes to 1 hour.

(Step 7b)

The olefin moiety of Compound [VIIi] may be oxidatively cleaved in asolvent to give Compound [VIIj].

The oxidative cleavage includes an ozone oxidation by a reductivetreatment.

A preferable solvent used in the reaction is a mixed solution ofchloroform and methanol.

The reaction temperature is usually about −100° C. to 0° C., preferablyabout −80° C. to 0° C.

The reaction time is usually about 5 minutes to 6 hours, preferablyabout 30 minutes to 2 hours.

The reagent used as the reducing agent includes dimethyl sulfide ortriphenylphosphine, preferably triphenylphosphine.

(Step 8b)

Compound [VIIj] may be reductively aminated in a solvent to giveCompound [VIIk].

A preferable amine used in the reaction is benzylamine.

A preferable solvent used in the reaction is tetrahydrofuran.

A preferable reaction temperature is room temperature.

A preferable reaction time is about 12 hours to 1 day.

A preferable reagent used as the reducing agent is sodiumtriacetoxyborohydride.

(Step 9b)

Compound [VIIk] may be treated by simultaneous removal of P^(C1) andP^(N5) in conventional carboxylic acid deprotection and aminedeprotection to give Compound [VIIl]. The deprotection may be carriedout by known methods depending on selected protecting groups.

For example, Compound [VIIk] wherein P^(C1) is tert-butyl ester andP^(N5) is tert-butoxycarbonyl may be treated in a single or a mixedsolvent of anisole, chloroform, ethyl acetate, dioxane, water, etc. byan acid such as hydrochloric acid, trifluoroacetic acid.

(Step 10b)

Compound [VIIl] may be intramolecularly cyclized in a solvent to giveCompound [VIIm]. The cyclization includes a conventional amidation inthe presence of a condensing agent and a base.

A preferable condensing agent used in the reaction isO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate.

A preferable base used in the reaction is diisopropylethylamine.

A preferable solvent used in the reaction is chloroform.

A preferable reaction temperature is room temperature.

The reaction time is usually about 30 minutes to 1 day, preferably about1 to 6 hours.

As an alternative cyclization, Compound [VIIk] wherein P^(C1) is ethylester or methyl ester may be selectively removed P^(N5) by aconventional amine deprotection with remaining P^(C1), followed by acyclization with 2M aqueous sodium hydroxide solution in an alcoholsolvent such as ethanol at room temperature to give Compound [VIIm].

(Step 11b)

The amide moiety of Compound [VIIm] may be reduced to an amine in asolvent, followed by an introduction of P^(N7) by a conventional amineprotection to give Compound [VIIn].

A preferable reducing agent used in the reduction is a mixture oflithium aluminum hydride and concentrated sulfuric acid. A preferableusage of concentrated sulfuric acid is 0.5 moles to 1 mole of lithiumaluminum hydride.

A preferable solvent used in the reduction is tetrahydrofuran.

The reaction temperature of the reduction is usually about 0° C. to roomtemperature, preferably 0° C.

The reaction time of the reduction is usually about 30 minutes to 3hours, preferably about 1 to 2 hours.

The amine protection may be carried out by known methods depending onselected protecting groups in the similar manner to the above GeneralPreparation Method 3 Step 1b. For example, in case that P^(N7) istert-butoxycarbonyl, Compound [VIIm] may be reacted with di-tert-butyldicarbonate in tetrahydrofuran.

(Step 12b)

P^(N6) among P^(N6) and P^(N7) of Compound [VIIn] may be selectivelyremoved by a conventional amine deprotection in the similar manner tothe above General Preparation Method 1 Step 2 to give Compound [VIIo].The deprotection may be carried out by known methods depending onselected protecting groups.

General Preparation Method 4

[In the above scheme, P^(N8) is a protecting group of amine, preferablytert-butoxycarbonyl; P^(C2) is a protecting group of carboxylic acid,preferably tert-butyl ester; Hal is a halogen atom; m2′ is 1 or 2;carbon atoms of * may be optionally substituted by R^(a) so as to bechemically acceptable; carbon atoms of # may be optionally substitutedby R^(b) so as to be chemically acceptable; R^(a), R^(b), n1, n2 and m1have the same meanings as defined in the above formula [I].]

(Step 1C)

Compound [VIIIa] may be reacted with trichloroethanal in a solvent togive Compound [VIIIb].

A preferable solvent used in the reaction is acetonitrile.

A preferable reaction temperature is room temperature.

The reaction time is usually about 30 minutes to 3 days, preferablyabout 6 hours to 1 day.

(Step 2c)

Compound [VIIIb] may be reacted with Compound [VIIIc] in a solvent inthe presence of a base to give Compound [VIIId].

A preferable solvent used in the reaction is tetrahydrofuran.

A preferable base used in the reaction is lithium diisopropylamide.

The reaction temperature is usually about −80° C. to room temperature,preferably about −80° C. to 0° C.

The reaction time is usually about 30 minutes to 1 day, preferably about1 to 3 hours.

(Step 3c)

Compound [VIIId] may be treated by solvolysis under an acidic conditionin an alcohol solvent to give Compound [VIIIe].

A preferable solvent used in the reaction is methanol.

A preferable acid used in the reaction is concentrated sulfuric acid. Apreferable usage of concentrated sulfuric acid is 0.1 to 3 moles to 1mole of Compound [VIIId].

A preferable reaction temperature is room temperature to 65° C.

The reaction time is usually about 30 minutes to 3 days, preferablyabout 6 hours to 1 day.

(Step 4c)

Compound [VIIIe] may be introduced P^(N8) by a conventional amineprotection in the similar manner to the above General Preparation Method3 Step 1b to give Compound [VIIIf]. The amine protection may be carriedout by known methods depending on selected protecting groups. Forexample, in case that P^(N8) is tert-butoxycarbonyl, Compound [VIIIe]may be reacted with di-tert-butyl dicarbonate in tetrahydrofuran.

(Step 5c)

Compound [VIIIf] may be treated in a solvent by a hydroboration,followed by an oxidation to give Compound [VIIIg] in the similar mannerto General Preparation Method 2 Step 4a-2.

(Step 6c)

Compound [VIIIg] may be oxidized in a solvent to give Compound [VIIIh].

The reagent used in the reaction includes sulfur trioxide-pyridinecomplex or Dess-Martin periodinane, preferably Dess-Martin periodinane.

A preferable solvent used in the reaction is chloroform.

A preferable reaction temperature is about 0° C. to room temperature.

The reaction time is usually about 30 minutes to 1 day, preferably about2 to 6 hours.

Bases including sodium bicarbonate as an additive may be optionallyadded.

The resulting Compound [VIIIh] may be reacted in the similar manner toGeneral Preparation Method 3 Step 8b or later to give an amine compoundcorresponding to Compound [VIIo].

EXAMPLES

Next, the preparations of the present compounds are specificallyillustrated by Examples. However, the present invention is not intendedto be limited thereto.

Stereochemistries in chemical structures of the compounds areabbreviated in the Examples.

Measurement apparatuses and conditions used in the Examples are asfollows.

HPLC analysis condition 1Preparation method for solution A: Sodium dihydrogen phosphate dihydrate(23.4 g) was dissolved in water (3000 mL) to be adjusted to pH 2.1 byusing phosphoric acid (10.2 mL).Measurement instrument: HPLC system SHIMADZU CORPORATION HighPerformance Liquid Chromatograph ProminenceColumn: DAICEL CHIRALPAK AD-3R 4.6 mmφ×150 mmColumn temperature: 40° C.Mobile phase: (solution A) 100 mM phosphate (sodium) buffer (pH 2.1),(solution B) methanol Solution A:Solution B=30:70 (constantly 20-minutesending).Sending rates of solutions: 0.5 ml/min

Detection: UV (220 nm) [Preparation 1]: Synthesis of Compound 1

(1) Piperidine-1,3-dicarboxylic acid 3-((E)-but-2-enyl)ester1-tert-butyl ester

To a solution of 1-(tert-butoxycarbonyl)-3-piperidine carboxylic acid(50.0 g) and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (45.8 g) inchloroform (500 ml) was added 4-dimethylaminopyridine (29.3 g), and themixture was stirred for 50 minutes. To the mixture was added(E)-but-2-en-1-ol (22.1 ml), and the mixture was stirred at roomtemperature for 1.5 hours. To the reaction mixture was added 10% aqueouspotassium bisulfate solution (500 ml), and the mixture was extractedwith chloroform. The separated organic layer was sequentially washedwith saturated aqueous sodium bicarbonate solution (400 ml) andsaturated aqueous sodium chloride solution (350 ml). The separatedaqueous layer was extracted with chloroform (300 ml) again. The combinedorganic layer was dried over anhydrous sodium sulfate, and concentratedunder reduced pressure. The resulting residue was purified by silica gelcolumn chromatography (eluent: n-hexane/ethyl acetate=7/1) to give thetitled compound (57.1 g).

¹H-NMR (CDCl₃) δ: 5.84-5.73 (1H, m), 5.62-5.52 (1H, m), 4.51 (2H, d,J=6.4 Hz), 4.37-4.00 (1H, m), 3.95-3.88 (1H, m), 3.19-2.87 (1H, m),2.84-2.76 (1H, m), 2.48-2.40 (1H, m), 2.08-2.01 (1H, m), 1.74-1.66 (5H,m), 1.63-1.55 (1H, m), 1.46 (9H, s).

(2) 3-(1-Methyl-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester

To a solution of piperidine-1,3-dicarboxylic acid3-((E)-but-2-enyl)ester 1-tert-butyl ester (105.7 g) in tetrahydrofuran(1000 ml) cooled to −68° C. was added lithium hexamethyl disilazide(1.1M tetrahydrofuran solution, 411 ml). The reaction mixture was warmedto 0° C. over 20 minutes, stirred at the same temperature for additional30 minutes, and then cooled to −68° C. again. To the mixture was addedtrimethylsilyl chloride (56.6 ml). The reaction mixture was warmed toroom temperature over 2 hours, and stirred at the same temperature foradditional 2 hours. After ice-cooling, to the mixture were sequentiallyadded water (1000 ml) and 2M aqueous sodium hydroxide solution (136 ml),and the mixture was washed with n-hexane (1000 ml). The separatedaqueous layer was acidified by 2M aqueous hydrochloric acid solution,and then extracted with ethyl acetate (800 ml). The organic layer wassequentially washed with water (700 ml) and saturated aqueous sodiumchloride solution (400 ml), and the separated aqueous layer wasextracted with ethyl acetate (600 ml) again. The combined organic layerwas dried over anhydrous sodium sulfate, and concentrated under reducedpressure. The resulting residue was slurry-washed with n-hexane/ethylacetate solution (6/1, 700 ml) to give the titled compound (59.8 g). Thefiltrate was concentrated under reduced pressure, and the resultingresidue was slurry-washed with n-hexane/ethyl acetate solution (4/1, 200ml) again to give a solid (14.4 g). The solid was slurry-washed withn-hexane/ethyl acetate solution (5/1, 100 ml) again to give the titledcompound (11.9 g). The resultant was combined to give the titledcompound (71.7 g).

¹H-NMR (CDCl₃) δ: 5.80-5.71 (1H, m), 5.09-5.07 (1H, m), 5.06-5.03 (1H,m), 4.32-4.20 (1H, m), 3.90-3.83 (1H, m), 2.87-2.74 (2H, m), 2.39-2.31(1H, m), 2.19-2.11 (1H, m), 1.64-1.55 (2H, m), 1.51-1.44 (1H, m), 1.45(9H, s), 1.07 (3H, d, J=7.1 Hz).

(3) Optically-Active Compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester

(3)-(1) Seed Crystal of a Salt of an Optically-Active Compound of3-(1-methyl-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butyl esterwith (R)-(−)-2-amino-2-phenyl-ethanol

3-(1-Methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester(2.0 g), isopropyl acetate (10 ml) and isopropanol (10 ml) were mixed todissolve. To the mixture was added (R)-(−)-2-amino-2-phenyl-ethanol (484mg), and the mixture was stirred at room temperature for 24 hours. Theslurry mixture was filtered, and the resulting solid was washed withisopropyl acetate (6 ml) and dried under reduced pressure to give thetitled compound (735 mg).

(3)-(2) Salt of an Optically-Active Compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl esterwith (R)-(−)-2-amino-2-phenyl-ethanol

3-(1-Methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester(106.6 g), (R)-(−)-2-amino-2-phenyl-ethanol (31.0 g), isopropyl acetate(480 ml) and isopropanol (480 ml) were mixed to dissolve at roomtemperature. To the mixture solution was added the seed crystal obtainedin (3)-(1), and the mixture was stirred for 16 hours. The slurry mixturewas filtered to give the titled compound (47.0 g). An analysis of theresulting solid by HPLC analysis condition 1 showed that an isomer withshorter retention times was a main product.

An isomer with shorter retention times (retention time 6.48 minutes)

An isomer with longer retention times (retention time 10.70 minutes)

(3)-(3) Optically-Active Compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl esterRecovered from the Filtrate of (3)-(2)

The filtrate of (3)-(2) was concentrated under reduced pressure, and theresulting residue was mixed with ethyl acetate (500 ml) and water (500ml). The mixture was acidified by the addition of potassium bisulfate.The separated organic layer was washed with saturated aqueous sodiumchloride solution and concentrated under reduced pressure. To theresidue was added isopropanol, and the mixture was concentrated underreduced pressure to give the titled compound (75.0 g).

¹H-NMR (CDCl₃) δ: 5.80-5.71 (1H, m), 5.09-5.07 (1H, m), 5.06-5.03 (1H,m), 4.32-4.20 (1H, m), 3.90-3.83 (1H, m), 2.87-2.74 (2H, m), 2.39-2.31(1H, m), 2.19-2.11 (1H, m), 1.64-1.55 (2H, m), 1.51-1.44 (1H, m), 1.45(9H, s), 1.07 (3H, d, J=7.1 Hz).

(4) Salt of an Optically-Active Compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl esterwith (S)-(+)-2-amino-2-phenyl-ethanol

The resulting residue (75.0 g) obtained in (3)-(3),(S)-(+)-2-amino-2-phenyl-ethanol (31.0 g), isopropyl acetate (335 ml)and isopropanol (306 ml) were mixed to dissolve at room temperature. Toa mixed solution was added the seed crystal obtained in (3)-(1), and themixture was stirred for 17.5 hours. The slurry mixture was filtered, andthe resulting solid was washed with isopropyl acetate (150 ml) to givethe titled compound (54.2 g). An analysis of the solid by HPLC analysiscondition 1 showed that an isomer with longer retention times was a mainproduct.

An isomer with shorter retention times (retention time 6.48 minutes)

An isomer with longer retention times (retention time 10.70 minutes)

(5) Optically-Active Compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester

To an optically-active compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester andsalt of (S)-(+)-2-amino-2-phenyl-ethanol (52.7 g) were added to mixethyl acetate (264 ml) and water (264 ml), and the mixture was acidifiedby the addition of potassium bisulfate. The separated organic layer waswashed with saturated aqueous sodium chloride solution, dried overanhydrous sodium sulfate, and concentrated under reduced pressure togive the titled compound (36.0 g). The crude product was partially usedin the next step without further purification.

¹H-NMR (CDCl₃) δ: 5.80-5.71 (1H, m), 5.09-5.07 (1H, m), 5.06-5.03 (1H,m), 4.32-4.20 (1H, m), 3.90-3.83 (1H, m), 2.87-2.74 (2H, m), 2.39-2.31(1H, m), 2.19-2.11 (1H, m), 1.64-1.55 (2H, m), 1.51-1.44 (1H, m), 1.45(9H, s), 1.07 (3H, d, J=7.1 Hz).

(6) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1-methylallyl)piperidine-1-carboxylic acidtert-butyl ester

To a refluxed solution of an optically-active compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester(9.0 g) and triethylamine (8.9 ml) in toluene (90 ml) was added dropwisediphenylphosphoryl azide (8.9 ml) over 25 minutes. The reaction mixturewas stirred for 2.5 hours at the same temperature, and then thereto wereadded benzyl alcohol (10 ml) and 4-dimethylaminopyridine (771 mg). Themixture was stirred for 33 hours with refluxing, followed by cooled toroom temperature, and acidified by the addition of 10% aqueous potassiumbisulfate solution. The mixture was extracted with ethyl acetate, andthe organic layer was washed with saturated aqueous sodium chloridesolution and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (eluent:chloroform/acetone=20/1). The fractions which could not be isolated orpurified were concentrated under reduced pressure, and the resultingresidue was purified by silica gel column chromatography (eluent:chloroform/acetone=20/1) again. The purified fractions are combined tobe concentrated under reduced pressure to give the titled compound (11.0g).

¹H-NMR (CDCl₃) δ: 7.37-7.29 (5H, m), 5.80-5.70 (1H, m), 5.36-5.17 (0.5H,m), 5.12-4.99 (4H, m), 4.96-4.74 (0.5H, m), 4.08-3.92 (2H, m), 2.98-2.33(4H, m), 1.64-1.38 (4H, m), 1.44 (9H, s), 1.03 (3H, d, J=7.0 Hz).

(7) Optically-Active Compound of3-benzyloxycarbonylamino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylicacid tert-butyl ester

A solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1-methylallyl)piperidine-1-carboxylic acid1-tert-butyl ester (25.5 g) in chloroform/methanol (250 ml/250 ml)cooled to −78° C. was flowed ozone air for 30 minutes. To the reactionmixture was added sodium borohydride (7.5 g) in small batches, and themixture was warmed to room temperature. To the mixture was added 5%aqueous sodium bicarbonate solution (250 ml), and the mixture wasextracted with chloroform (125 ml). The separated aqueous layer wasextracted with chloroform (125 ml) again. The combined organic layer wassequentially washed with a mixed aqueous solution of sodiumbicarbonate/sodium thiosulfate (12.5 g/25.0 g, 275 ml), water (125 ml)and 10% aqueous sodium chloride solution (125 ml), dried over anhydrousmagnesium sulfate, and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=1/0 to 2/3) to give the titled compound(21.9 g).

¹H-NMR (CDCl₃) δ: 7.37-7.29 (5H, m), 5.54-5.15 (1H, m), 5.06 (1H, d,J=12.4 Hz), 5.02 (1H, d, J=12.4 Hz), 4.03-3.93 (1H, m), 3.84-3.64 (3H,m), 3.27-2.84 (2H, m), 2.16-1.86 (2H, m), 1.66-1.59 (2H, m), 1.52-1.45(1H, m), 1.44 (9H, s), 1.04 (3H, d, J=7.1 Hz).

(8) Optically-Active Compound of3-amino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylic acidtert-butyl ester

To a solution of an optically-active compound of3-benzyloxycarbonylamino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylicacid tert-butyl ester (21.0 g) in methanol (400 ml) was added 10%palladium carbon (2.1 g), and the mixture was hydrogenated at roomtemperature under ordinary pressure for 90 minutes. The mixture wasfiltered through Celite, and the filtrate was concentrated under reducedpressure to give the titled compound (13.8 g).

¹H-NMR (CDCl₃) δ: 3.96-3.87 (1H, m), 3.86-3.77 (1H, m), 3.74-3.65 (1H,m), 3.63-3.58 (1H, m), 3.09-2.91 (1H, m), 2.87-2.77 (1H, m), 1.70-1.43(5H, m), 1.46 (9H, s), 0.96 (3H, d, J=6.9 Hz).

(9) Optically-Active Compound of3-methyl-1,6-diazaspiro[3,5]nonane-1,6-dicarboxylic acid 1-benzyl ester6-tert-butyl ester

To a solution of an optically-active compound of3-amino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylic acidtert-butyl ester (12.9 g), triphenylphosphine (20.3 g) and triethylamine(21.6 ml) in dichloromethane (400 ml) cooled to 0° C. was added carbontetrabromide (25.9 g) in small batches. The reaction mixture was stirredat room temperature for 1 hour, and the mixture was cooled to 4° C.Then, thereto were added triethylamine (10.8 ml) and benzylchloroformate (10.3 ml), and the mixture was stirred at room temperaturefor 40 minutes. To the mixture was added 5% aqueous sodium bicarbonatesolution (125 ml), and the mixture was extracted with chloroform (125ml). The separated aqueous layer was extracted with chloroform (125 ml)again. The combined organic layer was washed with water (125 ml), 10%aqueous sodium chloride solution (125 ml), dried over anhydrousmagnesium sulfate, and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=1/0 to 7/3) to give the titled compound(12.4 g).

¹H-NMR (CDCl₃) δ: 7.39-7.29 (5H, m), 5.17-5.00 (2H, m), 4.32-3.81 (3H,m), 3.48-3.02 (2H, m), 2.80-2.41 (2H, m), 2.24-2.14 (1H, m), 2.11-2.04(0.5H, m), 1.97-1.88 (0.5H, m), 1.70-1.39 (2H, m), 1.57 (9H, s), 1.18(3H, d, J=7.1 Hz).

(10) Optically-Active Compound of3-methyl-1,6-diazaspiro[3,5]nonane-1-carboxylic acid 1-benzyl ester

To a solution of an optically-active compound of3-methyl-1,6-diazaspiro[3,5]nonane-1,6-dicarboxylic acid 1-benzyl ester6-tert-butyl ester (5.54 g) in chloroform (83 ml) cooled to 4° C. wasadded a solution of trifluoroacetic acid/chloroform (28 ml/55 ml), andthe mixture was warmed to room temperature and stirred for additional 2hours. The reaction mixture was cooled to 4° C., and thereto was added4M aqueous sodium hydroxide solution (90 ml). The mixture was basifiedto be pH 9 to 10, and extracted with chloroform/methanol (4/1, 60 m1×2)twice. The combined organic layer was sequentially washed with 5%aqueous sodium bicarbonate solution, 10% aqueous sodium chloridesolution, dried over anhydrous magnesium sulfate, and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (eluent: chloroform/methanol/28% ammoniawater=90/10/1) to give the titled compound (2.20 g).

¹H-NMR (CDCl₃) δ: 7.40-7.29 (5H, m), 5.18-5.03 (2H, m), 4.02-3.94 (1H,m), 3.41-3.26 (1.5H, m), 3.15-3.08 (0.5H, m), 2.97-2.72 (2.5H, m),2.51-2.34 (1.5H, m), 2.24-2.13 (0.5H, m), 2.06-1.58 (3.5H, m), 1.53-1.35(1H, m), 1.30-1.15 (3H, m).

(11) Optically-Active Compound of3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester

An optically-active compound of3-methyl-1,6-diazaspiro[3,5]nonane-1-carboxylic acid 1-benzyl ester(2.20 g) was mixed with 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1.17 g),potassium carbonate (3.17 g) and water (12 ml), and the mixture wasstirred for 4 hours with refluxing. The mixture was cooled to roomtemperature, and extracted with chloroform twice. The combined organiclayer was washed with water, 10% aqueous sodium chloride solution, driedover anhydrous magnesium sulfate, and concentrated under reducedpressure. The resulting residue was purified by silica gel columnchromatography (eluent: chloroform/ethyl acetate=1/0 to 1/3, followed bychloroform/methanol=95/5) to give the titled compound (2.84 g).

¹H-NMR (CDCl₃) δ: 10.40-10.30 (1H, m), 8.32 (1H, s), 7.40-7.30 (5H, m),7.11-7.08 (1H, m), 6.57-6.51 (1H, m), 5.17-5.07 (2H, m), 5.04-4.89 (1H,m), 4.72-4.61 (1H, m), 4.14-4.07 (1H, m), 3.74-3.68 (0.5H, m), 3.57-3.51(0.5H, m), 3.39-3.32 (1H, m), 3.15-2.96 (1H, m), 2.57-2.47 (1H, m),2.44-2.32 (0.5H, m), 2.23-2.08 (1.5H, m), 1.92-1.80 (1H, m), 1.72-1.59(1H, m), 1.14-1.06 (3H, m).

(12) Optically-Active Compound of4-(3-methyl-1,6-diazaspiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine

To a solution of an optically-active compound of3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester (2.84 g) in methanol/tetrahydrofuran (14 ml/14 ml) wasadded 10% palladium carbon (568 mg), and the mixture was hydrogenatedunder 4 atmospheres. The mixture was filtered through Celite, and thefiltrate was concentrated under reduced pressure. The resulting residuewas slurry-washed with toluene/tetrahydrofuran (95/5, 9 ml) to give thetitled compound (1.74 g).

¹H-NMR (CDCl₃) δ: 10.63 (1H, br s), 8.32 (1H, s), 7.08 (1H, d, J=3.5Hz), 6.54 (1H, d, J=3.5 Hz), 4.45 (1H, d, J=13.0 Hz), 4.21-4.14 (1H, m),3.85-3.79 (1H, m), 3.57 (1H, d, J=13.0 Hz), 3.55-3.47 (1H, m), 3.08-3.01(1H, m), 2.48-2.38 (1H, m), 2.03 (1H, br s), 1.98-1.88 (1H, m),1.84-1.68 (3H, m), 1.19 (3H, d, J=7.3 Hz).

(13) Optically-Active Compound of3-[3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]non-1-yl]-3-oxopropionitrile

An optically-active compound of4-(3-methyl-1,6-diazaspiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine(1.8 g) was mixed with 1-cyanoacetyl-3,5-dimethylpyrazole (2.3 g),N,N-diisopropylethylamine (2.4 ml) and 1,4-dioxane (18 ml), and themixture was stirred at 100° C. for 3 hours. The mixture was cooled toroom temperature. Then, thereto were added water and saturated aqueoussodium chloride solution, and the mixture was extracted with ethylacetate. The separated aqueous layer was extracted with chloroform, andthe combined organic layer was concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: chloroform/ethyl acetate=1/1, followed bychloroform/methanol=20/1 to 10/1). The resulting residue was purified bysilica gel column chromatography (eluent: chloroform/acetone=10/1 to1/1) again. The resulting residue was purified by silica gel columnchromatography (eluent: chloroform/methanol=20/1 to 10/1) again to givethe titled compound (1.8 g).

¹H-NMR (DMSO-D₆) δ: 11.71 (1H, br s), 8.12 (1H, s), 7.20 (1H, dd, J=3.5,2.4 Hz), 6.65 (1H, dd, J=3.6, 1.9 Hz), 4.93-4.88 (1H, m), 4.64-4.58 (1H,m), 4.24-4.18 (1H, m), 3.67 (2H, s), 3.61 (1H, d, J=12.8 Hz), 3.46-3.41(1H, m), 3.03-2.95 (1H, m), 2.42-2.35 (1H, m), 2.34-2.25 (1H, m),2.15-2.08 (1H, m), 1.83-1.77 (1H, m), 1.57-1.43 (1H, m), 1.01 (3H, d,J=7.1 Hz).

[α]D=+168.100 (25° C., c=1.05, methanol)

[Preparation 2]: Synthesis of Compound 2

(1) Piperidine-1,3-dicarboxylic acid 3-((Z)-but-2-enyl)ester1-tert-butyl ester

To a solution of 1-(t-butoxycarbonyl)-3-piperidinecarboxylic acid (60.0g) and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (55.2 g) inchloroform (600 ml) was added 4-dimethylaminopyridine (35.2 g), and themixture was stirred for 70 minutes. To the mixture was added(Z)-but-2-en-1-ol (26.8 ml), and the mixture was stirred at roomtemperature for 14 hours. To the reaction mixture were added water (300ml) and saturated aqueous sodium chloride solution (100 ml), and themixture was extracted with chloroform. The separated organic layer wasconcentrated under reduced pressure, and to the resulting residue wereadded ethyl acetate and n-hexane. The mixture was sequentially washedwith 3.5% aqueous potassium bisulfate solution three times and saturatedaqueous sodium chloride solution twice. To the separated organic layerwas added silica gel (200 ml), and the mixture was stirred and filteredthrough Celite. The filtrate was concentrated under reduced pressure,and the resulting residue was purified by silica gel columnchromatography (eluent: n-hexane/ethyl acetate=9/1) to give the titledcompound (68.3 g).

¹H-NMR (DMSO-D₆) δ: 5.74-5.65 (1H, m), 5.55-5.47 (1H, m), 4.61 (2H, d,J=6.8 Hz), 4.05-3.75 (1H, m), 3.70-3.53 (1H, m), 2.96-2.86 (2H, m),2.46-2.39 (1H, m), 1.92-1.84 (1H, m), 1.69-1.51 (6H, m), 1.39 (9H, s).

(2) 3-(1-Methyl-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester

To a solution of piperidine-1,3-dicarboxylic acid3-((Z)-but-2-enyl)ester 1-tert-butyl ester (68.3 g) in tetrahydrofuran(700 ml) cooled to −74° C. was added lithium hexamethyl disilazide (1.6Mtetrahydrofuran solution, 166 ml). The reaction mixture was warmed to 0°C. over 35 minutes, stirred at the same temperature for additional 25minutes, and cooled to −74° C. again. To the mixture was addedtrimethylsilyl chloride (39.6 ml). The reaction mixture was warmed toroom temperature over 70 minutes, and stirred at the same temperaturefor additional 4 hours. To the mixture were sequentially added water(600 ml), 2M aqueous sodium hydroxide solution (70 ml), and the mixturewas washed with n-hexane (600 ml). The separated aqueous layer wasacidified by 2M aqueous hydrochloric acid solution, followed byextracted with ethyl acetate (300 ml, 200 ml, 150 ml) three times. Thecombined organic layer was washed with saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfate, and concentrated underreduced pressure. The resulting residue was slurry-washed withn-hexane/ethyl acetate solution (10/1) to give the titled compound (42.5g).

¹H-NMR (CDCl₃) δ: 5.83-5.72 (1H, m), 5.09-5.06 (1H, m), 5.05-5.02 (1H,m), 4.33-3.90 (1H, m), 3.73-3.63 (1H, m), 3.31-2.73 (2H, m), 2.43-2.34(1H, m), 2.07-1.97 (1H, m), 1.64-1.56 (2H, m), 1.54-1.42 (1H, m), 1.45(9H, s), 1.05 (3H, d, J=7.3 Hz).

(3) Salt of an Optically-Active Compound of3-(1-methyl-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butyl esterand (S)-(+)-2-amino-2-phenyl-ethanol

To a solution of 3-(1-methylallyl)piperidine-1,3-dicarboxylic acid1-tert-butyl ester (3.00 g) in 1,2-dimethoxyethane (30 ml) was added(S)-(+)-2-amino-2-phenyl-ethanol (800 mg), and the mixture was stirredat room temperature overnight. The slurry mixture was filtered to give asolid (1.8 g). To the solid was added 1,2-dimethoxyethane (45 ml), andthe mixture was dissolved at 80° C. The mixture was slurry-washed atroom temperature for 4 hours to give the titled compound (1.46 g). Ananalysis by HPLC analysis condition 1 showed that an isomer with longerretention times was a main product.

An isomer with shorter retention times (retention time 6.73 minutes)

An isomer with longer retention times (retention time 13.70 minutes)

(4) Optically-Active Compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester

To a salt of an optically-active compound of3-(1-methyl-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butyl esterand (S)-(+)-2-amino-2-phenyl-ethanol (1.5 g) were added ethyl acetate(15 ml) and water (15 ml), and the mixture was acidified by the additionof potassium bisulfate (567 mg). The separated aqueous layer wasextracted with ethyl acetate twice, and the combined organic layer waswashed with 10% aqueous potassium bisulfate solution twice and wateronce, dried over anhydrous magnesium sulfate, and concentrated underreduced pressure to give the titled compound (1.02 g).

¹H-NMR (CDCl₃) δ: 5.83-5.72 (1H, m), 5.09-5.06 (1H, m), 5.05-5.02 (1H,m), 4.33-3.90 (1H, m), 3.73-3.63 (1H, m), 3.31-2.73 (2H, m), 2.43-2.34(1H, m), 2.07-1.97 (1H, m), 1.64-1.56 (2H, m), 1.54-1.42 (1H, m), 1.45(9H, s), 1.05 (3H, d, J=7.3 Hz).

(5) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1-methylallyl)piperidine-1-carboxylic acidtert-butyl ester

To a solution of an optically-active compound of3-(1-methylallyl)piperidine-1,3-dicarboxylic acid 1-tert-butyl ester(1.02 g) and triethylamine (967 μl) in toluene (10 ml) heated to 90° C.was added dropwise diphenylphosphoryl azide (1.1 ml). The reactionmixture was stirred at the same temperature for 1 hour, and then theretowere added benzyl alcohol (718 μl) and 4-dimethylaminopyridine (127 mg).The mixture was stirred with refluxing overnight, then cooled to roomtemperature, and thereto were added water and ethyl acetate. Theseparated aqueous layer was extracted with ethyl acetate twice, and thecombined organic layer was washed with water twice and saturated aqueoussodium chloride solution once, dried over anhydrous magnesium sulfate,and concentrated under reduced pressure. The resulting residue waspurified by silica gel column chromatography (eluent:chloroform/acetone=50/1). The resulting residue was purified by silicagel column chromatography (eluent: n-hexane/ethyl acetate=20/1 to 4/1)to give the titled compound (845 mg).

¹H-NMR (DMSO-D₆) δ: 7.39-7.28 (5H, m), 6.91-6.72 (1H, m), 5.88-5.76 (1H,m), 5.18-4.84 (4H, m), 4.41-4.26 (1H, m), 3.88-3.61 (1H, m), 3.07-2.54(2H, m), 1.99-1.43 (3H, m), 1.35 (10H, s), 0.92-0.85 (3H, m).

(6) Optically-Active Compound of3-benzyloxycarbonylamino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylicacid tert-butyl ester

A solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1-methylallyl)piperidine-1-carboxylic acid1-tert-butyl ester (815 mg) in chloroform/methanol (6.6 ml/6.6 ml)cooled to −78° C. was flowed ozone air for 30 minutes. To the reactionmixture was added sodium borohydride (318 mg) in small batches, and thenthe mixture was warmed to room temperature over 40 minutes. To themixture was added saturated aqueous sodium bicarbonate solution, and themixture was extracted with chloroform twice. The combined organic layerwas washed with aqueous sodium bicarbonate solution, dried overanhydrous magnesium sulfate, and concentrated under reduced pressure.The resulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=3/2 to 0/1) to give the titled compound(406 mg).

¹H-NMR (DMSO-D₆) δ: 7.39-7.27 (5H, m), 6.74-6.55 (1H, m), 5.17-4.83 (2H,m), 4.53-4.41 (1H, m), 4.32-3.93 (2H, m), 3.76-3.47 (2H, m), 3.26-3.00(2H, m), 2.95-2.68 (1H, m), 2.30-2.00 (1H, m), 1.83-1.66 (1H, m),1.64-1.44 (2H, m), 1.36 (9H, s), 0.92-0.77 (3H, m).

(7) Optically-Active Compound of3-amino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylic acidtert-butyl ester

To a solution of an optically-active compound of3-benzyloxycarbonylamino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylicacid tert-butyl ester (374 mg) in methanol (6 ml) was added 10%palladium carbon (38 mg), and the mixture was hydrogenated at roomtemperature under ordinary pressure for 14 hours. The mixture wasfiltered through Celite, and the filtrate was concentrated under reducedpressure to give the titled compound (269 mg).

¹H-NMR (DMSO-D₆) δ: 4.83 (1H, br s), 3.51-2.74 (4H, m), 1.79-1.25 (7H,m), 1.38 (9H, s), 0.92 (3H, d, J=6.9 Hz).

(8) Optically-Active Compound of3-methyl-1,6-diazaspiro[3,5]nonane-1,6-dicarboxylic acid 1-benzyl ester6-tert-butyl ester

To a solution of an optically-active compound of3-amino-3-(2-hydroxy-1-methylethyl)piperidine-1-carboxylic acidtert-butyl ester (258 mg), triphenylphosphine (472 mg) and triethylamine(502 μl) in dichloromethane (7.7 ml) cooled to 0° C. was added carbontetrabromide (596 mg). The reaction mixture was stirred at roomtemperature for 2.5 hours and cooled to 4° C. Then, thereto were addedtriethylamine (279 μl) and benzyl chloroformate (267 μl), and themixture was stirred for 40 minutes. To the mixture was added water, andthe mixture was extracted with ethyl acetate. The separated organiclayer was sequentially washed with saturated aqueous sodium bicarbonatesolution and saturated aqueous sodium chloride solution, and dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=20/1 to 2/1) to give the titled compound(85 mg).

¹H-NMR (DMSO-D₆) δ: 7.40-7.29 (5H, m), 5.07-4.98 (2H, m), 4.20-4.11 (1H,m), 3.99-3.83 (2H, m), 3.39-3.20 (1H, m), 3.13-2.83 (1H, m), 2.58-2.45(1H, m), 2.44-2.32 (1H, m), 2.16-2.07 (0.5H, m), 2.01-1.82 (1.5H, m),1.68-1.58 (1H, m), 1.39 (9H, s), 1.37-1.28 (1H, m), 1.14-1.03 (3H, m).

(9) Optically-Active Compound of3-methyl-1,6-diazaspiro[3,5]nonane-1-carboxylic acid 1-benzyl ester

To a solution of an optically-active compound of3-methyl-1,6-diazaspiro[3,5]nonane-1,6-dicarboxylic acid 1-benzyl ester6-tert-butyl ester (78 mg) in chloroform (2 ml) cooled to 4° C. wasadded trifluoroacetic acid (0.4 ml), and the mixture was warmed to roomtemperature and stirred for additional 1 hour. The reaction mixture wasbasified by the addition of 1M aqueous sodium hydroxide solution, andextracted with chloroform. The separated organic layer was washed withsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfate and concentrated under reduced pressure to give the titledcompound (108 mg).

¹H-NMR (DMSO-D₆) δ: 7.42-7.31 (5.0H, m), 5.20-4.99 (2.0H, m), 4.05-3.88(1.0H, m), 3.64-3.55 (1.0H, m), 3.43-3.37 (0.5H, m), 3.35-3.26 (0.5H,m), 3.22-2.89 (3.0H, m), 2.39-2.29 (1.0H, m), 2.22-2.13 (0.5H, m),2.07-1.49 (4.0H, m), 1.29-1.22 (0.5H, m), 1.15 (1.5H, d, J=6.9 Hz),1.01-0.91 (0.5H, m).

(10) Optically-Active Compound of3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester

An optically-active compound of3-methyl-1,6-diazaspiro[3,5]nonane-1-carboxylic acid 1-benzyl ester (108mg) was mixed with 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (32 mg),potassium carbonate (86 mg) and water (2.1 ml), and stirred overnightwith refluxing. The mixture was cooled to room temperature, andextracted with chloroform. The separated organic layer was washed withsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (eluent: n-hexane/ethylacetate=2/1 to 0/1, followed by chloroform/methanol=9/1) to give thetitled compound (48 mg).

¹H-NMR (DMSO-D₆) δ: 11.69 (1.0H, br s), 8.15 (1.0H, s), 7.41-7.27 (5.0H,m), 7.18 (1.0H, dd, J=3.4, 2.6 Hz), 6.64-6.59 (1.0H, m), 5.09-4.92(3.5H, m), 4.63-4.55 (1.0H, m), 4.01-3.85 (1.0H, m), 3.45-3.39 (0.5H,m), 3.35-3.27 (0.5H, m), 3.25-3.20 (0.5H, m), 2.99-2.84 (1.0H, m),2.46-2.38 (1.0H, m), 2.35-2.25 (0.5H, m), 2.20-2.10 (0.5H, m), 2.03-1.95(1.0H, m), 1.88-1.79 (1.0H, m), 1.66-1.52 (1.0H, m), 0.93 (3.0H, d,J=6.9 Hz).

(11) Optically-Active Compound of4-(3-methyl-1,6-diazaspiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine

To a solution of an optically-active compound of3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester (46 mg) in methanol/tetrahydrofuran (1.8 ml/1.8 ml)was added 10% palladium carbon (15 mg), and the mixture was hydrogenatedunder 4 atmospheres for 14 hours. The mixture was filtered throughCelite, and the filtrate was concentrated under reduced pressure to givethe titled compound (27 mg).

¹H-NMR (DMSO-D₆) δ: 11.73 (1H, br s), 8.17 (1H, s), 7.21 (1H, dd, J=3.3,2.0 Hz), 6.64-6.61 (1H, m), 4.35 (0.5H, br s), 4.09 (0.5H, br s), 4.03(1H, d, J=13.5 Hz), 3.97 (1H, d, J=13.2 Hz), 3.83-3.70 (2H, m),3.47-3.40 (1H, m), 3.23-3.18 (1H, m), 2.48-2.43 (1H, m), 1.97-1.83 (2H,m), 1.77-1.57 (2H, m), 1.04 (3H, d, J=7.3 Hz).

(12) Optically-Active Compound of3-[3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]non-1-yl]-3-oxopropionitrile

An optically-active compound of4-(3-methyl-1,6-diazaspiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine (25mg) was mixed with 1-cyanoacetyl-3,5-dimethylpyrazole (32 mg) and1,4-dioxane (750 μl), and the mixture was stirred at 100° C. for 3.5hours. The mixture was cooled to room temperature. Thereto were addedwater and saturated aqueous sodium bicarbonate solution, and the mixturewas extracted with ethyl acetate. The separated aqueous layer wasextracted with ethyl acetate five times, and the combined organic layerwas washed with water and concentrated under reduced pressure. Theresulting residue was purified by silica-gel thin layer chromatography(eluent: chloroform/methanol=9/1). The resulting residue was purified bysilica-gel thin layer chromatography (eluent: ethylacetate/methanol=93/7) again. The resulting solid (8.0 mg) wasslurry-washed with n-hexane/ethyl acetate solution to give the titledcompound (6.7 mg).

¹H-NMR (DMSO-D6) δ: 11.70 (1H, br s), 8.14 (1H, s), 7.19 (1H, dd, J=3.5,2.4 Hz), 6.64 (1H, dd, J=3.6, 1.9 Hz), 5.03-4.96 (1H, m), 4.66-4.59 (1H,m), 4.11-4.06 (1H, m), 3.70 (1H, d, J=18.7 Hz), 3.65 (1H, d, J=18.7 Hz),3.60-3.55 (1H, m), 3.45 (1H, d, J=13.0 Hz), 2.97-2.89 (1H, m), 2.46-2.41(1H, m), 2.40-2.34 (1H, m), 2.00-1.94 (1H, m), 1.88-1.80 (1H, m),1.64-1.51 (1H, m), 0.91 (3H, d, J=7.1 Hz).

[α]D=+202.790 (25° C., c=1.04, methanol)

[Preparation 3]: Synthesis of Compound 3

(1) Piperidine-1,3-dicarboxylic acid 3-allyl ester 1-tert-butyl ester

To a solution of 1-(tert-butoxycarbonyl)-3-piperidinecarboxylic acid(50.0 g) in N,N-dimethylformamide (500 ml) were added potassiumcarbonate (60.3 g) and allyl bromide (28.3 ml), and the mixture wasstirred at room temperature for 3 hours. To the reaction mixture wasadded water (600 ml), and the mixture was extracted with ethyl acetate(600 ml). The organic layer was sequentially washed with water (600 ml)and saturated aqueous sodium chloride solution (400 ml). The separatedaqueous layer was extracted with ethyl acetate (300 ml) again. Thecombined organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. To a solution of the resultingresidue in n-hexane/ethyl acetate (4/1, 400 ml) was added silica gel (70g), and the mixture was stirred at room temperature. The mixture wasfiltered, and the filtrate was concentrated under reduced pressure togive the titled compound (62.5 g).

¹H-NMR (CDCl₃) δ: 5.97-5.85 (1H, m), 5.36-5.20 (2H, m), 4.63-4.55 (2H,m), 4.34-4.01 (1H, m), 3.97-3.86 (1H, m), 3.16-2.88 (1H, m), 2.86-2.77(1H, m), 2.53-2.43 (1H, m), 2.11-2.02 (1H, m), 1.75-1.57 (2H, m),1.52-1.39 (1H, m), 1.46 (9H, s).

(2) 3-Allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester

To a solution of piperidine-1,3-dicarboxylic acid 3-allyl ester1-tert-butyl ester (62.5 g) in tetrahydrofuran (625 ml) cooled to −72°C. was added lithium hexamethyl disilazide (1.6M tetrahydrofuransolution, 160 ml). The mixture was stirred at the same temperature for30 minutes, and then the reaction mixture was warmed to 0° C. over 12minutes and cooled to −67° C. again. To the mixture was addedtrimethylsilyl chloride (35.2 ml). The reaction mixture was warmed to2.5° C. over 1 hour, and stirred at the same temperature for additional2 hours. To the mixture were sequentially added methanol (250 ml) and 1Maqueous sodium hydroxide solution (250 ml), and the mixture was washedwith n-hexane (940 ml). The separated aqueous layer was washed withn-hexane (250 ml) again. The separated aqueous layer was acidified by 1Mhydrochloric acid water, and then extracted with ethyl acetate (800 ml).The organic layer was sequentially washed with water (800 ml) andsaturated aqueous sodium chloride solution (400 ml), and the separatedaqueous layer was extracted with ethyl acetate (500 ml) again. Thecombined organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The resulting residue wasslurry-washed with n-hexane/ethyl acetate (10/1, 660 ml) to give thetitled compound (53.4 g).

¹H-NMR (CDCl₃) δ: 5.81-5.69 (1H, m), 5.14-5.06 (2H, m), 3.93-3.78 (1H,m), 3.54-3.45 (1H, m), 3.29-3.14 (2H, m), 2.43-2.34 (1H, m), 2.29-2.21(1H, m), 2.07-1.98 (1H, m), 1.65-1.49 (3H, m), 1.45 (9H, s).

(3) Optically-Active Compound of 3-allylpiperidine-1,3-dicarboxylic acid1-tert-butyl ester

(3)-(1) Seed Crystal of a Salt of an Optically-Active Compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester with(S)-(−)-2-amino-3-phenyl-1-propanol

3-Allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester (3.0 g) wasmixed with isopropyl acetate (30 ml), and the mixture was heated to 80°C. to dissolve. To the mixture was added(S)-(−)-2-amino-3-phenyl-1-propanol (1.01 g), and the mixture wasstirred at room temperature for 21 hours. The slurry mixture wasfiltered, and the resulting solid was washed with isopropyl acetate (8ml) and dried under reduced pressure to give the titled compound (1.5g). The filtrate was concentrated under reduced pressure and used in(4)-(1).

(3)-(2) Salt of an Optically-Active Compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester with(S)-(−)-2-amino-3-phenyl-1-propanol

To a solution of 3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butylester (73.3 g) in isopropyl acetate (733 ml) heated to 80° C. was added(S)-(−)-2-amino-3-phenyl-1-propanol (24.7 g). The mixed solution wascooled to room temperature. Then, thereto was added the seed crystalobtained in (3)-(1), and the mixture was stirred overnight. The slurrymixture was filtered, and the resulting solid was washed with isopropylacetate (210 ml) to give the titled compound (37.4 g). An analysis byHPLC analysis condition 1 showed that an isomer with shorter retentiontimes was a main product.

An isomer with shorter retention times (retention time 6.01 minutes)

An isomer with longer retention times (retention time 8.94 minutes)

(3)-(3) Optically-Active Compound of 3-allylpiperidine-1,3-dicarboxylicacid 1-tert-butyl ester from the Filtrate of (3)-(2)

The filtrate of (3)-(2) was combined with the wash solution. Thereto wasadded aqueous potassium bisulfate solution (22.2 g/365 ml), and themixture was stirred. The separated organic layer was sequentially washedwith 10% aqueous potassium bisulfate solution, water and saturatedaqueous sodium chloride solution, and concentrated under reducedpressure to give the titled compound (49 g).

¹H-NMR (CDCl₃) δ: 5.81-5.69 (1H, m), 5.14-5.06 (2H, m), 3.93-3.78 (1H,m), 3.54-3.45 (1H, m), 3.29-3.14 (2H, m), 2.43-2.34 (1H, m), 2.29-2.21(1H, m), 2.07-1.98 (1H, m), 1.65-1.49 (3H, m), 1.45 (9H, s)

(4) Salt of an Optically-Active Compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester with(R)-(+)-2-amino-3-phenyl-1-propanol

(4)-(1) Seed Crystal of a Salt of an Optically-Active Compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester with(R)-(+)-2-amino-3-phenyl-1-propanol

To the residue (2.74 g) obtained from the filtrate of (3)-(1) were addedethyl acetate (14 ml) and water (14 ml), and the mixture was cooled to0° C. Then, thereto was added potassium bisulfate (494 mg). The mixturewas stirred at room temperature for 30 minutes, and then extracted withethyl acetate. The organic layer was sequentially washed with 10%aqueous potassium bisulfate solution, water (twice) and saturatedaqueous sodium chloride solution, dried over anhydrous magnesium sulfateand concentrated under reduced pressure. The resulting residue (1.94 g)was mixed with isopropyl acetate (20 ml), and the mixture was heated to80° C. to dissolve. To the mixture was added(R)-(+)-2-amino-3-phenyl-1-propanol (925 mg), and the mixture wasstirred at room temperature for 19.5 hours. The slurry mixture wasfiltered, and the resulting solid was dried under reduced pressure togive the titled compound (1.85 g).

(4)-(2) Salt of an Optically-Active Compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester with(R)-(+)-2-amino-3-phenyl-1-propanol

The residue (49 g) obtained in (3)-(3) and isopropyl acetate (490 ml)were combined, and the mixture was heated to 80° C. to dissolve. To themixture was added (R)-(+)-2-amino-3-phenyl-1-propanol (23.7 g), and themixture was cooled to room temperature. Then, thereto was added the seedcrystal obtained in (4)-(1), and the mixture was stirred overnight. Theslurry mixture was filtered and washed with isopropyl acetate (150 ml)to give the titled compound (52 g). An analysis by HPLC analysiscondition 1 showed that an isomer with longer retention times was a mainproduct.

An isomer with shorter retention times (retention time 6.01 minutes)

An isomer with longer retention times (retention time 8.94 minutes)

(5) Optically-Active Compound of 3-allylpiperidine-1,3-dicarboxylic acid1-tert-butyl ester

To a salt of an optically-active compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester with(R)-(+)-2-amino-3-phenyl-1-propanol (52 g) obtained in (4)-(2) wereadded ethyl acetate (260 ml) and aqueous potassium bisulfate solution(20.2 g/260 ml), and the mixture was stirred. The separated organiclayer was sequentially washed with 10% aqueous potassium bisulfatesolution, water and saturated aqueous sodium chloride solution. Theseparated aqueous layer was extracted with ethyl acetate (130 ml) again,and sequentially washed with water and saturated aqueous sodium chloridesolution. The combined organic layer was concentrated under reducedpressure to give the titled compound (32.2 g).

¹H-NMR (CDCl₃) δ: 5.81-5.69 (1H, m), 5.14-5.06 (2H, m), 3.93-3.78 (1H,m), 3.54-3.45 (1H, m), 3.29-3.14 (2H, m), 2.43-2.34 (1H, m), 2.29-2.21(1H, m), 2.07-1.98 (1H, m), 1.65-1.49 (3H, m), 1.45 (9H, s).

(6) Optically-Active Compound of3-allyl-3-benzyloxycarbonylaminopiperidine-1-carboxylic acid tert-butylester

To a solution of an optically-active compound of3-allylpiperidine-1,3-dicarboxylic acid 1-tert-butyl ester (32.2 g) andtriethylamine (33.2 ml) in toluene (320 ml) heated to 80° C. was addeddropwise diphenylphosphoryl azide (33.4 ml) over 50 minutes. Thereaction mixture was stirred at the same temperature for 2 hours andcooled to room temperature. To the mixture were added benzyl alcohol(24.6 ml) and 4-dimethylaminopyridine (2.9 g). The mixture was stirredat 100° C. for 21 hours and cooled to room temperature. Then, theretowere added ethanol (200 ml) and water (200 ml), and the mixture wasextracted with n-hexane (200 ml). The separated organic layer wassequentially washed with saturated aqueous ammonium chloride solution(200 ml) and saturated aqueous sodium chloride solution (150 ml), andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (eluent: n-hexane/ethyl acetate=1/0to 6/1) to give the titled compound (39.4 g).

¹H-NMR (DMSO-D₆) δ: 7.39-7.27 (5H, m), 6.90-6.68 (1H, m), 5.75-5.63 (1H,m), 5.13-4.85 (4H, m), 4.07-3.94 (0.5H, m), 3.63-3.47 (1H, m), 3.39-3.12(1H, m), 3.11-2.88 (1.5H, m), 2.79-2.54 (1H, m), 2.28-2.01 (1H, m),1.79-1.66 (1H, m), 1.63-1.45 (3H, m), 1.36 (9H, s).

(7) Optically-Active Compound of3-benzyloxycarbonylamino-3-(2-hydroxyethyl)piperidine-1-carboxylic acidtert-butyl ester

A solution of an optically-active compound3-allyl-3-benzyloxycarbonylaminopiperidine-1-carboxylic acid tert-butylester (39.4 g) in chloroform/methanol (493 ml/493 ml) cooled to −78° C.was flowed ozone air for 1 hour. To the reaction mixture was addedsodium borohydride (19.9 g) in small batches, and the mixture was warmedto 4° C. over 50 minutes. To the mixture were added saturated aqueoussodium bicarbonate solution (200 ml) and water (400 ml), and the mixturewas extracted with chloroform (200 ml). The separated aqueous layer wasextracted with chloroform (300 ml) again, and the combined organic layerwas concentrated under reduced pressure. The resulting residue wasslurry-washed with n-hexane/ethyl acetate (1/1, 394 ml) solution to givea solid (23.5 g). The filtrate was concentrated under reduced pressure,and the resulting residue was slurry-washed with n-hexane/ethyl acetate(1/1) solution again to give a solid (2.4 g). Then, the filtrate wasconcentrated under reduced pressure, and the resulting residue waspurified by silica gel column chromatography (eluent: n-hexane/ethylacetate=1/1 to 0/1) to give a solid (1.5 g). The solids were combined togive the titled compound (27.4 g).

¹H-NMR (DMSO-D₆) δ: 7.39-7.26 (5H, m), 6.90-6.62 (1H, m), 5.12-4.87 (2H,m), 4.36 (1H, t, J=4.9 Hz), 4.00-2.96 (6H, m), 2.15-1.72 (2H, m),1.72-1.46 (2H, m), 1.46-1.17 (2H, m), 1.36 (9H, s).

(8) Optically-Active Compound of3-amino-3-(2-hydroxyethyl)piperidine-1-carboxylic acid tert-butyl ester

To a solution of an optically-active compound of3-benzyloxycarbonylamino-3-(2-hydroxyethyl)piperidine-1-carboxylic acidtert-butyl ester (27.2 g) in methanol (544 ml) was added 10% palladiumcarbon (2.7 g), and the mixture was hydrogenated under ordinary pressurefor 5 hours. The mixture was filtered through Celite, and the filtratewas concentrated under reduced pressure to give the titled compound(17.2 g).

¹H-NMR (DMSO-D₆) δ: 3.58-3.53 (2H, m), 3.43-2.85 (5H, m), 1.63-1.24 (6H,m), 1.39 (9H, s).

(9) Optically-Active Compound of1,6-diazaspiro[3,5]-nonane-1,6-dicarboxylic acid 1-benzyl ester6-tert-butyl ester

To a solution of an optically-active compound of3-amino-3-(2-hydroxyethyl)piperidine-1-carboxylic acid tert-butyl ester(6.46 g), triphenylphosphine (12.5 g) and triethylamine (13.3 ml) indichloromethane (226 ml) cooled to 0° C. was added carbon tetrabromide(15.8 g). The reaction mixture was stirred at room temperature for 40minutes and cooled to 0° C. again. Then, thereto were addedtriethylamine (5.5 ml) and benzyl chloroformate (4.9 ml), and themixture was stirred for 3.5 hours at room temperature. To the mixturewas added water (200 ml), and the mixture was extracted with chloroform(100 ml). The organic layer was washed with saturated aqueous sodiumchloride solution, and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=5/1) to give the titled compound (5.35g).

¹H-NMR (DMSO-D₆) δ: 7.40-7.29 (5H, m), 5.07-4.98 (2H, m), 4.10-3.96 (1H,m), 3.91-3.69 (3H, m), 2.64-2.53 (1H, m), 2.13-1.83 (4H, m), 1.66-1.54(1H, m), 1.43-1.27 (2H, m), 1.39 (9H, s).

(10) Optically-Active Compound of 1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester

To a solution of an optically-active compound of1,6-diazaspiro[3,5]-nonane-1,6-dicarboxylic acid 1-benzyl ester6-tert-butyl ester (5.35 g) in chloroform (134 ml) cooled to 0° C. wasadded trifluoroacetic acid (27 ml), and the mixture was stirred for 30minutes, warmed to room temperature and stirred for additional 30minutes. The reaction mixture was cooled to 0° C., and thereto was added4M aqueous sodium hydroxide solution (91 ml). The aqueous layer wasextracted with chloroform (100 ml, 50 ml) twice, and the combinedorganic layer was washed with saturated aqueous sodium chloride solutionand concentrated under reduced pressure. The resulting residue waspurified by silica gel column chromatography (eluent:chloroform/methanol=20/1 to 4/1) to give the titled compound (2.37 g).

¹H-NMR (DMSO-D₆) δ: 7.41-7.28 (5H, m), 5.09-4.96 (2H, m), 3.83-3.76 (1H,m), 3.73-3.66 (1H, m), 2.94-2.69 (3H, m), 2.38-2.15 (2H, m), 2.12-1.77(4H, m), 1.58-1.45 (1H, m), 1.38-1.26 (1H, m).

(11) Optically-Active Compound of6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester

An optically-active compound of 1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester (2.37 g) was mixed with4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1.4 g), potassium carbonate (3.8g) and water (71 ml), and heated to 100° C. to stir for 3 hours. Themixture was cooled to room temperature. Then, thereto was added waterand the mixture was extracted with ethyl acetate. The organic layer waswashed with saturated aqueous sodium chloride solution and concentratedunder reduced pressure. The resulting residue was slurry-washed withn-hexane/ethyl acetate (1/1) solution to give the titled compound (2.55g).

¹H-NMR (DMSO-D₆) δ: 11.72 (1H, s), 8.13 (1H, s), 7.41-7.28 (5H, m), 7.19(1H, dd, J=3.5, 2.6 Hz), 6.64-6.60 (1H, m), 5.13-5.02 (2H, m), 4.96-4.89(1H, m), 4.64-4.56 (1H, m), 3.98-3.70 (2H, m), 3.46-3.28 (1H, m),3.01-2.82 (1H, m), 2.31-2.22 (0.5H, m), 2.16-2.06 (0.5H, m), 2.03-1.90(3H, m), 1.83-1.75 (1H, m), 1.61-1.47 (1H, m).

(12) Optically-Active Compound of4-(1,6-diazaspiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine

To a solution of an optically-active compound of6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]nonane-1-carboxylicacid benzyl ester (2.55 g) in methanol/tetrahydrofuran (51 ml/51 ml) wasadded 20% palladium carbon (510 mg), and the mixture was hydrogenatedunder 4 atmospheres for 5 hours. The mixture was filtered throughCelite, and the filtrate was concentrated under reduced pressure. Theresulting residue was slurry-washed with diisopropyl ether (30 ml) togive the titled compound (1.55 g).

¹H-NMR (DMSO-D₆) δ: 11.68 (1H, br s), 8.12 (1H, s), 7.18 (1H, d, J=2.6Hz), 6.61 (1H, d, J=3.2 Hz), 4.18 (1H, d, J=12.8 Hz), 4.00-3.93 (1H, m),3.59 (1H, d, J=12.8 Hz), 3.54-3.47 (1H, m), 3.42-3.16 (3H, m), 2.03-1.82(3H, m), 1.72-1.62 (2H, m), 1.57-1.46 (1H, m).

(13) Optically-Active Compound of3-oxo-3-[6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.5]non-1-yl]-propionitrile

An optically-active compound of4-(1,6-diazaspiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine (1.55 g) wasmixed with 1-cyanoacetyl-3,5-dimethylpyrazole (1.56 g),N,N-diisopropylethylamine (1.66 ml) and 1,4-dioxane (31 ml), and themixture was heated to 100° C. to stir for 2 hours. The mixture wascooled to room temperature. Then, thereto was added saturated aqueoussodium bicarbonate solution, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated aqueous sodiumchloride solution, and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: ethyl acetate/methanol=20/1 to 9/1) to give the titled compound(1.50 g). 1H-NMR (DMSO-D6) δ: 11.73 (1H, br s), 8.13 (1H, s), 7.22-7.18(1H, m), 6.67-6.63 (1H, m), 4.96 (1H, d, J=12.6 Hz), 4.68-4.60 (1H, m),4.11-4.03 (1H, m), 4.01-3.93 (1H, m), 3.71 (2H, s), 3.53 (1H, d, J=12.6Hz), 2.98-2.88 (1H, m), 2.40-2.30 (1H, m), 2.03-1.90 (3H, m), 1.83-1.75(1H, m), 1.59-1.45 (1H, m).

[α]D=+210.000 (25° C., c=1.01, methanol)

[Preparation 4]: Synthesis of Compound 4

(1) Pyrrolidine-1,3-dicarboxylic acid 1-tert-butyl ester3-(3,3-difluoroallyl)ester

To a solution of pyrrolidine-1,3-dicarboxylic acid-1-tert-butyl ester(3.40 g) in N,N-dimethylformamide (34 ml) were added potassium carbonate(4.37 g) and 3-bromo-3,3-difluoropropene (1.93 ml), and the mixture wasstirred at 60° C. for 6 hours. To the reaction mixture was added water,and the mixture was extracted with ethyl acetate. The organic layer wasdried over anhydrous sodium sulfate, and concentrated under reducedpressure. The resulting residue was purified by silica gel columnchromatography (eluent: n-hexane/ethyl acetate=5/1) to give the titledcompound (3.73 g).

¹H-NMR (CDCl₃) δ: 4.64-4.48 (3H, m), 3.67-3.42 (3H, m), 3.41-3.29 (1H,m), 3.10-2.98 (1H, m), 2.18-2.04 (2H, m), 1.46 (9H, s).

(2) 3-(1,1-Difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester

To a solution of pyrrolidine-1,3-dicarboxylic acid 1-tert-butyl ester3-(3,3-difluoroallyl)ester (3.73 g) in tetrahydrofuran (37 ml) cooled to−78° C. was added lithium hexamethyl disilazide (1.0M tetrahydrofuransolution, 14 ml). The reaction mixture was warmed to 0° C. over 4minutes, stirred at the same temperature for additional 14 minutes, andthen cooled to −78° C. again. To the mixture was added trimethylsilylchloride (2.0 ml). The reaction mixture was warmed to room temperature,and stirred at the same temperature for additional 3 hours. The mixturewas basified by the addition of water, followed by 2M aqueous sodiumhydroxide solution to pH 8 to 9, and washed with n-hexane/diethylether(4/3). To the separated organic layer was added n-hexane, and themixture was extracted with 2M aqueous sodium hydroxide solution. Thecombined aqueous layer was washed with n-hexane/diethylether (4/3) 7times, acidified by 10% aqueous citric acid solution, and extracted withethyl acetate. The organic layer was concentrated under reducedpressure, and the resulting residue was slurry-washed withn-hexane/ethyl acetate (5/1) solution to give the titled compound (2.00g).

¹H-NMR (DMSO-D₆) δ: 13.47 (1H, br s), 6.23-6.09 (1H, m), 5.71-5.64 (1H,m), 5.63-5.58 (1H, m), 3.79-3.72 (1H, m), 3.47-3.28 (2H, m), 3.26-3.15(1H, m), 2.36-2.25 (1H, m), 2.21-2.06 (1H, m), 1.39 (9H, s).

(3) Optically-Active Compound of3-(1,1-difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester

An optically-active compound of3-(1,1-difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester (1.5 g), (S)-(−)-1-(1-naphthyl)ethylamine (826 μl) and isopropylacetate (15 ml) were combined, and the mixture was stirred at roomtemperature for 21 hours. The slurry mixture was filtered to remove asolid. The filtrate was concentrated under reduced pressure, and theresulting residue was mixed with ethyl acetate (20 ml) and 10% aqueouspotassium bisulfate solution (20 ml). The mixture was stirred at 0° C.for 1 hour. The separated organic layer was sequentially washed withwater and saturated aqueous sodium chloride solution, dried overanhydrous magnesium sulfate, and concentrated with under reducedpressure to give the titled compound (869 mg).

¹H-NMR (DMSO-D₆) δ: 13.47 (1H, br s), 6.23-6.09 (1H, m), 5.71-5.64 (1H,m), 5.63-5.58 (1H, m), 3.79-3.72 (1H, m), 3.47-3.28 (2H, m), 3.26-3.15(1H, m), 2.36-2.25 (1H, m), 2.21-2.06 (1H, m), 1.39 (9H, s).

(4) Salt of an Optically-Active Compound of3-(1,1-difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester with (R)-(+)-1-(1-naphthyl)ethylamine

The residue (869 mg) obtained in (3) was mixed with(R)-(+)-1-(1-naphthyl)ethylamine (263 μl) and isopropyl acetate (13 ml),and the mixture was stirred at room temperature overnight. The slurrymixture was filtered, and the resulting solid was washed with isopropylacetate (3 ml) to give the titled compound (754 mg). Analysis of thesolid by HPLC analysis condition 1 showed that an isomer with longerretention times was a main product.

An isomer with shorter retention times (retention time 5.32 minutes)

An isomer with longer retention times (retention time 7.01 minutes)

(5) Optically-Active Compound of3-(1,1-difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester

A salt of an optically-active compound of3-(1,1-difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester with (R)-(+)-1-(1-naphthyl)ethylamine (741 mg) was mixed withethyl acetate (7.4 ml) and water (3.7 ml), and the mixture was acidifiedby the addition of potassium bisulfate (261 mg). The separated organiclayer was washed with saturated aqueous sodium chloride solution, andconcentrated under reduced pressure to give the titled compound (457mg).

¹H-NMR (DMSO-D₆) δ: 13.47 (1H, br s), 6.23-6.09 (1H, m), 5.71-5.64 (1H,m), 5.63-5.58 (1H, m), 3.79-3.72 (1H, m), 3.47-3.28 (2H, m), 3.26-3.15(1H, m), 2.36-2.25 (1H, m), 2.21-2.06 (1H, m), 1.39 (9H, s).

(6) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1,1-difluoroallyl)pyrrolidine-1-carboxylicacid tert-butyl ester

To a solution of an optically-active compound of3-(1,1-difluoroallyl)pyrrolidine-1,3-dicarboxylic acid 1-tert-butylester (10.6 g) and triethylamine (10.2 ml) in toluene (106 ml) heated to100° C. was added dropwise diphenylphosphoryl azide (10.2 ml). Thereaction mixture was stirred at 100° C. for 50 minutes, and then theretowere added benzyl alcohol (5.6 ml) and 4-dimethylaminopyridine (0.89 g).The mixture was stirred at 100° C. for 16.5 hours, and then cooled toroom temperature. Then, thereto was added 5% aqueous potassium bisulfatesolution, and the mixture was extracted with toluene. The organic layerwas sequentially washed with 1M aqueous sodium hydroxide solution andsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (eluent:n-hexane/chloroform/ethyl acetate=5/4/1 to 4/4/1) to give the titledcompound (15.5 g).

¹H-NMR (CDCl₃) δ: 7.40-7.30 (5.0H, m), 6.04-5.89 (1.0H, m), 5.76-5.68(1.0H, m), 5.56-5.49 (1.0H, m), 5.07 (2.0H, s), 4.90-4.83 (1.0H, m),3.85-3.79 (0.5H, m), 3.74-3.40 (3.5H, m), 2.90-2.76 (0.5H, m), 2.63-2.49(0.5H, m), 2.27-2.18 (1.0H, m), 1.46 (9.0H, s).

(7) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-hydroxyethyl)pyrrolidine-1-carboxylicacid tert-butyl ester

A solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1,1-difluoroallyl)pyrrolidine-1-carboxylicacid tert-butyl ester (15.5 g) in chloroform/methanol (154 ml/154 ml)cooled to −78° C. was flowed ozone air for 30 minutes. To the reactionmixture was added sodium borohydride (4.14 g) in small batches, and thenthe mixture was warmed to 0° C. and stirred at the same temperature foradditional 4 hours. To the mixture were added saturated aqueous sodiumbicarbonate solution and saturated aqueous sodium chloride solution, andthe mixture was extracted with chloroform. The separated aqueous layerwas extracted with ethyl acetate. The combined organic layer was driedover anhydrous sodium sulfate, and concentrated under reduced pressure.The resulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=3/1). The fractions which could not beseparated and isolated were concentrated under reduced pressure, and theresulting residue was purified by silica gel column chromatography(eluent: n-hexane/chloroform/ethyl acetate=5/4/1) again. The purifiedfractions were combined and concentrated under reduced pressure to givethe titled compound (12.8 g).

¹H-NMR (CDCl₃) δ: 7.42-7.32 (5.0H, m), 5.15-5.07 (2.0H, m), 5.04-4.93(1.0H, m), 4.02-3.49 (6.0H, m), 3.43-3.30 (1.0H, m), 2.67-2.57 (0.4H,m), 2.45-2.28 (1.6H, m), 1.46 (9.0H, s).

(8) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-methanesulfonyloxyethyl)pyrrolidine-1-carboxylicacid tert-butyl ester

To a solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-hydroxyethyl)pyrrolidine-1-carboxylicacid tert-butyl ester (471 mg) and triethylamine (328 μl) in chloroform(4.7 ml) cooled to 0° C. was added methanesulfonyl chloride (118 μl),and the mixture was stirred for 50 minutes. To the mixture was added 5%aqueous potassium bisulfate solution, and the mixture was extracted withethyl acetate. The organic layer was sequentially washed with 1M aqueoussodium hydroxide solution and saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfate and concentrated underreduced pressure to give the titled compound (565 mg).

¹H-NMR (CDCl₃) δ: 7.41-7.33 (5H, m), 5.10 (2H, s), 4.94-4.86 (1H, m),4.57-4.48 (2H, m), 3.92-3.84 (0.5H, m), 3.80-3.52 (2.5H, m), 3.47-3.33(1H, m), 3.09 (3H, s), 2.83-2.74 (0.5H, m), 2.61-2.52 (0.5H, m),2.31-2.21 (1H, m), 1.47 (9H, s).

(9) Optically-Active Compound of3,3-difluoro-1,6-diazaspiro[3,4]octane-1,6-dicarboxylic acid 1-benzylester 6-tert-butyl ester

To a solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-methanesulfonyloxyethyl)pyrrolidine-1-carboxylicacid tert-butyl ester (565 mg) in N,N-dimethylformamide (17 ml) cooledto 0° C. was added sodium hydride (71 mg, with 40% mineral oil), and themixture was stirred at the same temperature for 1 hour. To the reactionmixture was added water, and the mixture was extracted with ethylacetate. The organic layer was washed with saturated aqueous sodiumchloride solution, dried over anhydrous sodium sulfate and concentratedunder reduced pressure to give the titled compound (397 mg).

¹H-NMR (CDCl₃) δ: 7.41-7.31 (5H, m), 5.25-5.06 (2H, m), 4.27-4.17 (2H,m), 3.96-3.82 (1H, m), 3.79-3.49 (2H, m), 3.41-3.27 (1H, m), 2.65-2.41(0.5H, m), 2.38-2.17 (1.5H, m), 1.50-1.44 (9H, m).

(10) Optically-Active Compound of3,3-difluoro-1,6-diazaspiro[3,4]octane-6-carboxylic acid tert-butylester

To a solution of an optically-active compound of3,3-difluoro-1,6-diazaspiro[3,4]octane-1,6-dicarboxylic acid 1-benzylester 6-tert-butyl ester (397 mg) in methanol/tetrahydrofuran (3.2ml/3.2 ml) was added 10% palladium carbon (79 mg), and the mixture washydrogenated at room temperature under ordinary pressure for 14 hours.The mixture was filtered through Celite, and the filtrate wasconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (eluent: chloroform/acetone=4/1 to2/1). The fractions which could not be separated and isolated wereconcentrated under reduced pressure, and the resulting residue waspurified by silica gel column chromatography (eluent:chloroform/acetone=2/1 to 1/1) again. The purified fractions werecombined and concentrated under reduced pressure to give the titledcompound (211 mg).

¹H-NMR (CDCl₃) δ: 3.96-3.74 (3H, m), 3.55-3.37 (2H, m), 3.35-3.25 (1H,m), 2.45-2.35 (1H, m), 2.00-1.88 (1H, m), 1.73-1.56 (1H, m), 1.46 (9H,s).

(11) 3,3-Difluoro-1,6-diazaspiro[3,4]octane 2 hydrochloride

To an optically-active compound of3,3-difluoro-1,6-diazaspiro[3,4]octane-6-carboxylic acid tert-butylester (211 mg) were added 4M hydrochloric acid-1,4-dioxane (2.1 ml) and2M hydrochloric acid-methanol (2.1 ml), and the mixture was stirred atroom temperature for 45 minutes. To the reaction mixture was added ethylacetate, and the mixture was filtered to give the titled compound (163mg).

¹H-NMR (DMSO-D₆) δ: 9.80-9.42 (2H, m), 4.58-3.83 (7H, m), 3.47-3.33 (2H,m), 2.82-2.72 (1H, m).

(12) Optically-Active Compound of4-(3,3-difluoro-1,6-diazaspiro[3,4]oct-6-yl)-7H-pyrrolo[2,3-d]pyrimidine

An optically-active compound of 3,3-difluoro-1,6-diazaspiro[3,4]octane 2hydrochloride (163 mg) was mixed with4-chloro-7H-pyrrolo[2,3-d]pyrimidine (113 mg), potassium carbonate (509mg) and water (2.8 ml), and the mixture was stirred for 16.5 hours withrefluxing. The mixture was cooled to room temperature. Thereto was addedwater, and the mixture was filtered to give a solid (126 mg). Thefiltrate was extracted with ethyl acetate/methanol 4 times andchloroform/methanol twice. The combined organic layer was dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresulting residue was combined with the solid and purified by silica gelcolumn chromatography (eluent: chloroform/methanol=92/8 to 85/15) togive the titled compound (170 mg).

¹H-NMR (DMSO-D₆) δ: 11.63 (1H, br s), 8.10 (1H, s), 7.14 (1H, dd, J=3.6,2.4 Hz), 6.57 (1H, dd, J=3.6, 2.0 Hz), 4.15 (1H, d, J=11.7 Hz),3.92-3.65 (5H, m), 3.35-3.22 (1H, m), 2.47-2.39 (1H, m), 2.17-2.07 (1H,m).

(13) Optically-Active Compound of3-[3,3-difluoro-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.4]oct-1-yl]-3-oxopropionitrile

An optically-active compound of4-(3,3-difluoro-1,6-diazaspiro[3,4]oct-6-yl)-7H-pyrrolo[2,3-d]pyrimidine(170 mg) was mixed with 1-cyanoacetyl-3,5-dimethylpyrazole (209 mg),N,N-diisopropylethylamine (117 μl) and 1,4-dioxane (3.4 ml), and themixture was stirred at 100° C. for 75 minutes. The mixture was cooled toroom temperature. Thereto was added saturated aqueous sodium bicarbonatesolution, and the mixture was extracted with ethyl acetate. Theseparated aqueous layer was extracted with ethyl acetate twice again.The combined organic layer was washed with saturated aqueous sodiumchloride solution, dried over anhydrous sodium sulfate and concentratedunder reduced pressure. The resulting residue was purified by silica gelcolumn chromatography (eluent: chloroform/ethyl acetate=2/1, followed bychloroform/methanol=92/8 to 3/1). The mixture was concentrated underreduced pressure, and the resulting residue was slurry-washed with amixed solvent of n-heptane/ethanol (3/1) to give the titled compound(177 mg).

¹H-NMR (DMSO-D₆) δ: 11.68 (1H, br s), 8.11 (1H, s), 7.17-7.14 (1H, m),6.58 (1H, dd, J=3.3, 1.8 Hz), 4.66-4.58 (2H, m), 4.24-4.13 (2H, m),4.12-4.02 (1H, m), 3.89-3.78 (3H, m), 2.68-2.58 (1H, m), 2.56-2.45 (1H,m).

[α]D=+46.67° (25° C., c=0.54, methanol)

[Preparation 5]: Synthesis of Compound 5

(1) Piperidine-1,3-dicarboxylic acid 1-tert-butyl ester3-(3,3-difluoro-allyl)ester

To a solution of piperidine-1,3-dicarboxylic acid 1-tert-butyl ester(30.0 g) in N,N-dimethylformamide (300 ml) were added potassiumcarbonate (36.2 g) and 3-bromo-3,3-difluoropropene (16 ml), and themixture was stirred at 60° C. for 4.5 hours. To the reaction mixture wasadded water, and the mixture was extracted with n-hexane/ethyl acetate(1/1) solution. The organic layer was dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (eluent: n-hexane/ethylacetate=10/1) to give the titled compound (33.5 g).

¹H-NMR (CDCl₃) δ: 4.62-4.46 (3H, m), 4.34-3.98 (1H, m), 3.95-3.80 (1H,m), 3.18-2.89 (1H, m), 2.87-2.77 (1H, m), 2.50-2.40 (1H, m), 2.08-1.98(1H, m), 1.75-1.55 (2H, m), 1.52-1.41 (1H, m), 1.46 (9H, s).

(2) 3-(1,1-Difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester

To a solution of piperidine-1,3-dicarboxylic acid 1-tert-butyl ester3-(3,3-difluoro-allyl)ester (28.5 g) in tetrahydrofuran (285 ml) cooledto −78° C. was added lithium hexamethyl disilazide (1.1M tetrahydrofuransolution, 110 ml). The reaction mixture was warmed to −2° C. over 6minutes, stirred at the same temperature for additional 15 minutes, andthen cooled to −78° C. again. To the mixture was added trimethylsilylchloride (19 ml). The reaction mixture was warmed to room temperature,and stirred at the same temperature for additional 30 minutes. Themixture was basified by the addition of water (300 ml), followed by 2Maqueous sodium hydroxide solution (60 ml), and washed with n-hexane (450ml). The separated organic layer was extracted with 2M aqueous sodiumhydroxide solution 3 times. The combined aqueous layer was acidified by10% aqueous citric acid solution and extracted with ethyl acetate. Theorganic layer was concentrated under reduced pressure, and the resultingresidue was slurry-washed with n-hexane/ethyl acetate (5/1) solution togive the titled compound (23.9 g).

¹H-NMR (CDCl₃) δ: 6.05-5.89 (1H, m), 5.72-5.65 (1H, m), 5.56-5.52 (1H,m), 4.67 (1H, d, J=13.7 Hz), 4.12-4.02 (1H, m), 3.46 (1H, br s), 2.92(1H, d, J=13.7 Hz), 2.70-2.61 (1H, m), 2.37-2.30 (1H, m), 1.71-1.54 (3H,m), 1.45 (9H, s).

(3) Optically-Active Compound of3-(1,1-difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester

An optically-active compound of3-(1,1-difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester (26.9 g), (R)-(−)-2-amino-2-phenyl-ethanol (6.6 g) and isopropanol(270 ml) were mixed, and the mixture was stirred at room temperatureovernight. The slurry mixture was filtered to remove a solid. Thefiltrate was concentrated under reduced pressure, and the resultingresidue was mixed with ethyl acetate (97 ml) and 25% aqueous potassiumbisulfate solution (97 ml), and the mixture was stirred for 25 minutes.The separated organic layer was sequentially washed with 5% aqueouspotassium bisulfate solution and saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfate and concentrated underreduced pressure to give the titled compound (16.2 g).

¹H-NMR (CDCl₃) δ: 6.05-5.89 (1H, m), 5.72-5.65 (1H, m), 5.56-5.52 (1H,m), 4.67 (1H, d, J=13.7 Hz), 4.12-4.02 (1H, m), 3.46 (1H, br s), 2.92(1H, d, J=13.7 Hz), 2.70-2.61 (1H, m), 2.37-2.30 (1H, m), 1.71-1.54 (3H,m), 1.45 (9H, s).

(4) Salt of an Optically-Active Compound of3-(1,1-difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester with (S)-(+)-2-amino-2-phenyl-ethanol

The residue (16.2 g) obtained in (3) was mixed with(S)-(+)-2-amino-2-phenyl-ethanol (6.6 g) and isopropanol (162 ml), andthe mixture was stirred at room temperature for 20 hours. The slurrymixture was filtered, and the resulting solid was washed withisopropanol (80 ml) to give a solid (17.8 g). The solid was mixed withisopropanol (305 ml), and stirred at 105° C. for 20 minutes and at roomtemperature for 19 hours. The slurry mixture was filtered, and theresulting solid was washed with isopropanol (90 ml) to give the titledcompound (16.0 g). An analysis by HPLC analysis condition 1 showed thatan isomer with longer retention times was a main product.

An isomer with shorter retention times (retention time 5.41 minutes)

An isomer with longer retention times (retention time 12.47 minutes)

(5) Optically-Active Compound of3-(1,1-difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester

To a salt of an optically-active compound of3-(1,1-difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester with (S)-(+)-2-amino-2-phenyl-ethanol (17.8 g) were added ethylacetate (89 ml) and 25% aqueous potassium bisulfate solution (89 ml),and the mixture was stirred for 15 minutes. The separated organic layerwas sequentially washed with 5% aqueous potassium bisulfate solution (89ml) and saturated aqueous sodium chloride solution, dried over anhydroussodium sulfate and concentrated under reduced pressure to give thetitled compound (12.3 g).

¹H-NMR (CDCl₃) δ: 6.05-5.89 (1H, m), 5.72-5.65 (1H, m), 5.56-5.52 (1H,m), 4.67 (1H, d, J=13.7 Hz), 4.12-4.02 (1H, m), 3.46 (1H, br s), 2.92(1H, d, J=13.7 Hz), 2.70-2.61 (1H, m), 2.37-2.30 (1H, m), 1.71-1.54 (3H,m), 1.45 (9H, s).

(6) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-allyl)-piperidine-1-carboxylicacid tert-butyl ester

To a solution of an optically-active compound of3-(1,1-difluoro-allyl)-piperidine-1,3-dicarboxylic acid 1-tert-butylester (12.3 g) and triethylamine (11.2 ml) in toluene (123 ml) heated to100° C. was added dropwise diphenylphosphoryl azide (11.2 ml). Thereaction mixture was stirred at 100° C. for 25 minutes, and then theretowere added benzyl alcohol (6.2 ml) and 4-dimethylaminopyridine (0.98 g).The mixture was stirred at 100° C. overnight, and then cooled to roomtemperature and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography (eluent:n-hexane/ethyl acetate=9/1 to 85/15) to give the titled compound (15.1g).

¹H-NMR (CDCl₃) δ: 7.38-7.28 (5H, m), 6.07-5.92 (1H, m), 5.71-5.62 (1H,m), 5.53-5.48 (1H, m), 5.36 (0.5H, br s), 5.09-4.97 (2H, m), 4.89 (0.5H,br s), 4.28-3.98 (2H, m), 3.16-2.61 (3H, m), 1.64-1.52 (3H, m),1.49-1.36 (9H, m).

(7) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-hydroxy-ethyl)-piperidine-1-carboxylicacid tert-butyl ester

A solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-allyl)-piperidine-1-carboxylicacid tert-butyl ester (15.1 g) in chloroform/methanol (150 ml/150 ml)cooled to −78° C. was flowed ozone air for 30 minutes. To the reactionmixture was added sodium borohydride (4.2 g) in small batches, and thenthe mixture was warmed to 0° C. and stirred at the same temperature foradditional 50 minutes. To the mixture were added saturated aqueoussodium bicarbonate solution and saturated aqueous sodium chloridesolution, and the mixture was extracted with chloroform. The separatedaqueous layer was extracted with ethyl acetate. The combined organiclayer was dried over anhydrous sodium sulfate, and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (eluent: n-hexane/ethyl acetate=3/1 to 2/1) togive the titled compound (13.6 g).

¹H-NMR (CDCl₃) δ: 7.38-7.29 (5H, m), 5.78 (1H, br s), 5.15-4.99 (2H, m),4.42-3.97 (2H, m), 3.95-3.75 (2H, m), 3.55-3.44 (1H, m), 3.15-2.60 (3H,m), 1.79-1.67 (1H, m), 1.55-1.50 (1H, m), 1.42 (9H, s).

(8) Optically-Active Compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-methanesulfonyloxy-ethyl)-piperidine-1-carboxylicacid tert-butyl ester

To a solution of an optically-active compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-hydroxy-ethyl)-piperidine-1-carboxylicacid tert-butyl ester (13.6 g) and triethylamine (11.4 ml) in chloroform(136 ml) cooled to 0° C. was added methanesulfonyl chloride (7.8 ml),and the mixture was stirred for 50 minutes. To the mixture was added 5%aqueous potassium bisulfate solution, and the mixture was extracted withethyl acetate. The separated organic layer was sequentially washed with4M aqueous sodium hydroxide solution twice and saturated aqueous sodiumchloride solution once, dried over anhydrous sodium sulfate, andconcentrated under reduced pressure to give the titled compound (17.1g).

¹H-NMR (CDCl₃) δ: 7.39-7.29 (5H, m), 5.65 (1H, br s), 5.15-4.97 (2H, m),4.65-4.51 (2H, m), 4.39-3.98 (2H, m), 3.15-2.62 (3H, m), 3.07 (3H, s),1.70-1.53 (3H, m), 1.43 (9H, s).

(9) Optically-Active Compound of3,3-difluoro-1,6-diaza-spiro[3.5]nonane-1,6-dicarboxylic acid 1-benzylester 6-tert-butyl ester

To a suspension of sodium hydride (1.7 g, with 40% mineral oil) inN,N-dimethylformamide (428 ml) cooled to 0° C. was added a solution ofan optically-active compound of3-benzyloxycarbonylamino-3-(1,1-difluoro-2-methanesulfonyloxy-ethyl)-piperidine-1-carboxylicacid tert-butyl ester (17.1 g) in N,N-dimethylformamide (86 ml), and themixture was stirred at the same temperature for 15 minutes. To thereaction mixture was added water, and the mixture was extracted withethyl acetate. The separated organic layer was sequentially washed withwater and saturated aqueous sodium chloride solution, dried overanhydrous sodium sulfate and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=5/1) to give the titled compound (12.6g).

¹H-NMR (CDCl₃) δ: 7.41-7.30 (5H, m), 5.21-5.02 (2H, m), 4.56-4.31 (1H,m), 4.25-4.14 (2H, m), 4.13-3.89 (1H, m), 3.41-2.98 (1H, m), 2.80-2.41(1H, m), 2.33-2.12 (1H, m), 2.03-1.87 (1H, m), 1.80-1.67 (1H, m),1.65-1.52 (2H, m), 1.45 (9H, s).

(10) Optically-Active Compound of3,3-difluoro-1,6-diaza-spiro[3.5]nonane-1-carboxylic acid benzyl ester 1hydrochloride

To an optically-active compound of3,3-difluoro-1,6-diaza-spiro[3.5]nonane-1,6-dicarboxylic acid 1-benzylester 6-tert-butyl ester (12.6 g) was added 4M hydrochloricacid-1,4-dioxane (126 ml), and the mixture was stirred at roomtemperature for 30 minutes. The mixture was concentrated under reducedpressure, and thereto was added toluene. The mixture was concentratedunder reduced pressure again to give the titled compound (11.2 g).

¹H-NMR (CDCl₃) δ: 11.56 (1H, br s), 8.65 (1H, br s), 7.43-7.32 (5H, m),5.13 (2H, s), 4.43-4.20 (2H, m), 3.82-3.72 (1H, m), 3.54-3.35 (2H, m),3.28-3.14 (1H, m), 2.34-1.91 (4H, m).

(11) Optically-Active Compound of3,3-difluoro-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diaza-spiro[3.5]nonane-1-carboxylicacid benzyl ester

An optically-active compound of3,3-difluoro-1,6-diaza-spiro[3.5]nonane-1-carboxylic acid benzyl ester 1hydrochloride (11.2 g) was mixed with4-chloro-7H-pyrrolo[2,3-d]pyrimidine (4.2 g), potassium carbonate (18.7g) and water (104 ml), and the mixture was stirred for 16.5 hours withrefluxing. The mixture was cooled to room temperature, and thereto wereadded ethyl acetate and water. The mixture was filtered, and thefiltrate was separated and the resulting organic layer was washed withsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (eluent:chloroform/acetone=3/1). The resulting residue was purified by silicagel column chromatography (eluent: chloroform/acetone=2/1) again to givethe titled compound (12.0 g).

¹H-NMR (CDCl₃) δ: 10.22 (1H, br s), 8.37 (1H, s), 7.42-7.31 (5H, m),7.10 (1H, dd, J=3.3, 2.2 Hz), 6.56-6.51 (1H, m), 5.25-5.06 (3H, m),4.77-4.62 (1H, m), 4.34-4.18 (2H, m), 3.75-3.43 (1H, m), 3.14-2.94 (1H,m), 2.52-2.38 (0.5H, m), 2.36-2.27 (1H, m), 2.23-2.11 (0.5H, m),2.04-1.91 (1H, m), 1.90-1.77 (1H, m).

(12) Optically-Active Compound of4-(3,3-difluoro-1,6-diaza-spiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine

To a solution of an optically-active compound of3,3-difluoro-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diaza-spiro[3.5]nonane-1-carboxylicacid benzyl ester (12.0 g) in methanol/tetrahydrofuran (120 ml/120 ml)was added 10% palladium carbon (2.4 g), and the mixture was hydrogenatedat room temperature under 4 atmospheres for 21 hours. The mixture wasfiltered through Celite, and the filtrate was concentrated under reducedpressure. The resulting residue was slurry-purified by a mixed solventof n-hexane/ethyl acetate (2/1) to give the titled compound (6.6 g).

¹H-NMR (CDCl₃) δ: 10.04 (1H, br s), 8.32 (1H, s), 7.10 (1H, d, J=3.5Hz), 6.53 (1H, d, J=3.7 Hz), 4.60 (1H, d, J=13.7 Hz), 4.20-4.13 (1H, m),4.08-3.97 (1H, m), 3.92-3.81 (1H, m), 3.76 (1H, d, J=13.7 Hz), 3.64-3.55(1H, m), 2.08-1.78 (5H, m).

(13) Optically-Active Compound of3-[3,3-difluoro-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diaza-spiro[3.5]non-1-yl]-3-oxo-propionitrile

An optically-active compound of4-(3,3-difluoro-1,6-diaza-spiro[3.5]non-6-yl)-7H-pyrrolo[2,3-d]pyrimidine(6.6 g) was mixed with 1-cyanoacetyl-3,5-dimethylpyrazole (7.7 g),N,N-diisopropylethylamine (4.3 ml) and 1,4-dioxane (132 ml), and themixture was stirred at 100° C. for 2 hours. The mixture was cooled toroom temperature. Then, thereto was added water, and the mixture wasextracted with ethyl acetate. The separated aqueous layer was furtherextracted with ethyl acetate 5 times. The combined organic layer wassequentially washed with saturated aqueous sodium bicarbonate solutionand saturated aqueous sodium chloride solution, dried over anhydroussodium sulfate and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography (eluent:chloroform/acetone=3/2 to 2/3). To the resulting residue was addedn-heptane, and the mixture was concentrated under reduced pressure. Tothe resulting residue was added diethylether, and the mixture wasslurry-washed and filtered to give a solid (7.5 g). The filtrate wasconcentrated under reduced pressure. To the resulting residue was addeddiethylether, and the mixture was slurry-washed to give a solid (0.2 g).The combined solid was purified by silica gel column chromatography(eluent: chloroform/methanol=97/3 to 9/1). To the resulting solid wasadded ethanol, and the mixture was concentrated under reduced pressureto give the titled compound (7.1 g).

¹H-NMR (DMSO-D6) δ: 11.74 (1H, br s), 8.15 (1H, s), 7.22-7.19 (1H, m),6.65-6.62 (1H, m), 5.13-5.07 (1H, m), 4.69-4.46 (3H, m), 3.83 (1H, d,J=18.8 Hz), 3.77 (1H, d, J=19.2 Hz), 3.61-3.55 (1H, m), 2.99-2.91 (1H,m), 2.39-2.29 (1H, m), 2.26-2.19 (1H, m), 1.99-1.91 (1H, m), 1.63-1.48(1H, m).

[α]D=+139.52° (25° C., c=1.04, methanol)

[Preparation 6]: Synthesis of Compound 6

(1) Optically-Active Compound of 2-benzylaminopropan-1-ol

To a solution of (S)-(+)-2-aminopropan-1-ol (50.0 g) and benzaldehyde(74 ml) in ethanol (500 ml) was added 5% palladium carbon (5.0 g), andthe mixture was hydrogenated at room temperature under ordinary pressurefor 8 hours. The reaction mixture was filtered through Celite, andconcentrated under reduced pressure to give the titled compound (111.2g).

¹H-NMR (DMSO-D₆) δ: 7.34-7.27 (4H, m), 7.23-7.18 (1H, m), 4.53-4.47 (1H,m), 3.76 (1H, d, J=13.5 Hz), 3.66 (1H, d, J=13.5 Hz), 3.29-3.24 (2H, m),2.65-2.55 (1H, m), 1.99 (1H, br s), 0.93 (3H, d, J=6.4 Hz).

(2) Optically-Active Compound of[benzyl-(2-hydroxy-1-methylethyl)-amino]acetic acid tert-butyl ester

To a mixture of an optically-active compound of 2-benzylaminopropan-1-ol(111.2 g), potassium carbonate (111.6 g) and N,N-dimethylformamide (556ml) cooled to 0° C. was added dropwise bromoacetic acid tert-butyl ester(109 ml) over 20 minutes, and the mixture was stirred at roomtemperature for 19.5 hours. The mixture was acidified by the addition of2M aqueous hydrochloric acid solution and 6M aqueous hydrochloric acidsolution to pH 2, and washed with toluene (1000 ml). The separatedorganic layer was extracted with 0.1M aqueous hydrochloric acid solution(300 ml). The combined aqueous layer was adjusted to pH 10 by 4M aqueoussodium hydroxide solution, and extracted with ethyl acetate (700 ml).The organic layer was sequentially washed with water (900 ml) andsaturated aqueous sodium chloride solution (500 ml). The separatedaqueous layer was extracted with ethyl acetate (400 ml) again. Thecombined organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure to give the titled compound (160.0g).

¹H-NMR (DMSO-D₆) δ: 7.37-7.26 (4H, m), 7.24-7.19 (1H, m), 4.26 (1H, dd,J=6.9, 3.9 Hz), 3.76 (1H, d, J=14.1 Hz), 3.68 (1H, d, J=13.9 Hz),3.45-3.39 (1H, m), 3.29-3.20 (1H, m), 3.24 (1H, d, J=17.2 Hz), 3.13 (1H,d, J=17.0 Hz), 2.84-2.74 (1H, m), 1.37 (9H, s), 0.96 (3H, d, J=6.8 Hz).

(3) Optically-Active Compound of [benzyl-(2-chloropropyl)-amino]aceticacid tert-butyl ester

(3)-(1) Optically-Active Compound of[benzyl-(2-chloro-1-methylethyl)-amino]acetic acid tert-butyl ester

To a solution of an optically-active compound of[benzyl-(2-hydroxy-1-methylethyl)-amino]acetic acid tert-butyl ester(160.0 g) in chloroform (640 ml) cooled to 0° C. was added dropwisethionyl chloride (50.0 ml), and the mixture was stirred at 60° C. for 2hours. The reaction mixture was cooled to 0° C. Then, thereto were addedsaturated aqueous sodium bicarbonate solution (1000 ml) and chloroform(100 ml), and the mixture was stirred. The separated organic layer waswashed with saturated aqueous sodium chloride solution (500 ml), and theaqueous layer was extracted with chloroform (450 ml) again. The combinedorganic layer was dried over anhydrous sodium sulfate and concentratedunder reduced pressure to give the titled compound (172.9 g).

¹H-NMR (CDCl₃) δ: 7.40-7.22 (5H, m), 4.05-3.97 (0.4H, m), 3.93-3.81 (2H,m), 3.70-3.65 (0.6H, m), 3.44-3.38 (0.6H, m), 3.29 (0.8H, s), 3.27(1.2H, d, J=2.4 Hz), 3.24-3.15 (0.6H, m), 3.05-2.99 (0.4H, m), 2.94-2.88(0.4H, m), 1.50 (1.2H, d, J=6.4 Hz), 1.48 (3.6H, s), 1.45 (5.4H, s),1.23 (1.8H, d, J=6.8 Hz).

(3)-(2) Optically-Active Compound of[benzyl-(2-chloropropyl)-amino]acetic acid tert-butyl ester

An optically-active compound of[benzyl-(2-chloro-1-methylethyl)-amino]acetic acid tert-butyl ester(172.9 g) was dissolved in N,N-dimethylformamide (520 ml), and stirredat 80° C. for 140 minutes. The reaction mixture was cooled to 0° C.Then, thereto was added water (1200 ml), and the mixture was extractedwith n-hexane/ethyl acetate (2/1, 1000 ml). The organic layer wassequentially washed with water (700 ml) and saturated aqueous sodiumchloride solution (400 ml), and the separated aqueous layer wasextracted with n-hexane/ethyl acetate (2/1, 600 ml) again. The combinedorganic layer was concentrated under reduced pressure, and the resultingresidue was purified by silica gel column chromatography (eluent:n-hexane/ethyl acetate=50/1 to 40/1) to give the titled compound (127.0g).

¹H-NMR (CDCl₃) δ: 7.37-7.29 (4H, m), 7.28-7.23 (1H, m), 4.05-3.97 (1H,m), 3.91 (1H, d, J=13.5 Hz), 3.86 (1H, d, J=13.7 Hz), 3.29 (2H, s), 3.03(1H, dd, J=13.9, 6.6 Hz), 2.91 (1H, dd, J=13.9, 6.8 Hz), 1.50 (3H, d,J=6.4 Hz), 1.48 (9H, s).

(4) Optically-Active Compound of 1-benzyl-3-methylazetidine-2-carboxylicacid tert-butyl ester

To a solution of an optically-active compound of[benzyl-(2-chloropropyl)-amino]acetic acid tert-butyl ester (60.0 g) andhexamethyl phosphoramide (36.0 ml) in tetrahydrofuran (360 ml) cooled to−72° C. was added dropwise lithium hexamethyl disilazide (1.0Mtetrahydrofuran solution, 242 ml) over 18 minutes, and the mixture waswarmed to 0° C. over 80 minutes. To the reaction mixture weresequentially added saturated aqueous ammonium chloride solution (300 ml)and water (400 ml), and the mixture was extracted with ethyl acetate(500 ml). The organic layer was sequentially washed with water (700 ml)and saturated aqueous sodium chloride solution (500 ml), and theseparated aqueous layer was extracted with ethyl acetate (300 ml) again.The combined organic layer was dried over anhydrous sodium sulfate, andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (eluent: n-hexane/ethyl acetate=50/1to 4/1) to give the titled compound (50.9 g).

¹H-NMR (CDCl₃) δ: 7.34-7.21 (5H, m), 3.75 (1H, d, J=12.6 Hz), 3.70-3.67(1H, m), 3.58 (1H, d, J=12.6 Hz), 3.05-3.01 (1H, m), 2.99-2.95 (1H, m),2.70-2.59 (1H, m), 1.41 (9H, s), 1.24 (3H, d, J=7.1 Hz).

(5) Optically-Active Compound of 3-methylazetidine-1,2-dicarboxylic aciddi-tert-butyl ester

To a solution of an optically-active compound of1-benzyl-3-methylazetidine-2-carboxylic acid tert-butyl ester (43.5 g)and di-tert-butyl dicarbonate (38.2 g) in tetrahydrofuran/methanol (130ml/130 ml) was added 20% palladium hydroxide carbon (3.5 g), and themixture was hydrogenated under 4 atmospheres for 2 hours. The mixturewas filtered through Celite, and the filtrate was concentrated underreduced pressure to give the titled compound (48.0 g).

¹H-NMR (DMSO-D₆) δ: 4.44 (1H, d, J=8.8 Hz), 3.99-3.77 (1H, m), 3.45-3.37(1H, m), 3.00-2.88 (1H, m), 1.45 (9H, s), 1.40-1.30 (9H, m), 1.02 (3H,d, J=7.2 Hz).

(6) Optically-Active Compound of3-methyl-2-(3-methyl-but-2-enyl)-azetidine-1,2-dicarboxylic aciddi-tert-butyl ester

To a solution of an optically-active compound of3-methylazetidine-1,2-dicarboxylic acid di-tert-butyl ester (48.0 g) and1-bromo-3-methyl-2-butene (25.4 ml) in tetrahydrofuran (380 ml) cooledto −69° C. was added lithium hexamethyl disilazide (1.0M tetrahydrofuransolution, 200 ml). The reaction mixture was warmed to −20° C. over 40minutes, and stirred at the same temperature for additional 20 minutes.To the reaction mixture were sequentially added saturated aqueousammonium chloride solution (200 ml) and water (300 ml), and the mixturewas extracted with n-hexane/ethyl acetate (1/1, 500 ml). The separatedorganic layer was sequentially washed with water (200 ml) and saturatedaqueous sodium chloride solution (200 ml), dried over anhydrousmagnesium sulfate and concentrated under reduced pressure. The resultingresidue was purified by silica gel column chromatography (eluent:n-hexane/ethyl acetate=15/1 to 8/1) to give the titled compound (44.5g).

¹H-NMR (CDCl₃) δ: 5.29-5.21 (1H, m), 3.77-3.72 (1H, m), 3.49-3.44 (1H,m), 2.73-2.52 (3H, m), 1.76-1.74 (3H, m), 1.66-1.65 (3H, m), 1.51 (9H,s), 1.43 (9H, s), 1.05 (3H, d, J=7.3 Hz).

(7) Optically-Active Compound of3-methyl-2-(2-oxoethyl)azetidine-1,2-dicarboxylic acid di-tert-butylester

A solution of an optically-active compound of3-methyl-2-(3-methyl-but-2-enyl)-azetidine-1,2-dicarboxylic aciddi-tert-butyl ester (44.5 g) in chloroform/methanol (310 ml/310 ml)cooled to −70° C. was flowed ozone air for 1 hour. To the reactionmixture was added a solution of triphenylphosphine (44.7 g) inchloroform (45 ml) in small batches, and the mixture was warmed to roomtemperature. To the mixture were added saturated aqueous sodiumthiosulfate solution (200 ml) and water (300 ml), and the mixture wasextracted with chloroform (500 ml). The separated organic layer waswashed with saturated aqueous sodium chloride solution, dried overanhydrous magnesium sulfate and concentrated under reduced pressure togive the titled compound (95.0 g), which was used in the next stepwithout further purification.

¹H-NMR (DMSO-D₆) δ: 9.65 (1H, t, J=2.6 Hz), 3.79-3.74 (1H, m), 3.45-3.40(1H, m), 2.99-2.80 (3H, m), 1.46 (9H, s), 1.34 (9H, s), 1.06 (3H, d,J=7.2 Hz).

(8) Optically-Active Compound of2-(2-benzylaminoethyl)-3-methylazetidine-1,2-dicarboxylic aciddi-tert-butyl ester

To a solution of the residue (95.0 g) obtained in (7) in tetrahydrofuran(300 ml) was added benzylamine (34 ml) at room temperature, and themixture was stirred for 2 hours. The mixture was cooled to 0° C. Then,thereto was added sodium triacetoxyborohydride (83.3 g), and the mixturewas stirred at room temperature for 1.5 hours. To the reaction mixturewas added water (300 ml), and the mixture was extracted withn-hexane/ethyl acetate (1/3, 600 ml). The separated organic layer waswashed with water (300 ml) and saturated aqueous sodium chloridesolution (200 ml), and then extracted with κ% aqueous citric acidsolution (300 ml, 200 ml) twice and 10% aqueous citric acid solution(250 ml×3) 3 times. The combined aqueous layer was basified by 4Maqueous sodium hydroxide solution to pH 10 and extracted with chloroform(300 ml). The organic layer was washed with saturated aqueous sodiumchloride solution (200 ml), dried over anhydrous magnesium sulfate andconcentrated under reduced pressure to give the titled compound (46.9g).

¹H-NMR (DMSO-D₆) δ: 7.34-7.26 (4H, m), 7.22-7.17 (1H, m), 3.74-3.65 (2H,m), 3.61 (1H, t, J=7.8 Hz), 3.28 (1H, t, J=7.5 Hz), 2.76-2.66 (2H, m),2.57-2.45 (1H, m), 2.15 (1H, br s), 2.05-1.89 (2H, m), 1.42 (9H, s),1.27 (9H, s), 0.96 (3H, d, J=7.1 Hz).

(9) Optically-Active Compound of2-(2-benzylaminoethyl)-3-methylazetidine-2-dicarboxylic acid 2hydrochloride

An optically-active compound of2-(2-benzylaminoethyl)-3-methylazetidine-1,2-dicarboxylic aciddi-tert-butyl ester (46.5 g) was mixed with 4M hydrochloricacid-1,4-dioxane (230 ml) and water (4.1 ml), and the mixture wasstirred at 80° C. for 2 hours. The mixture was concentrated underreduced pressure, azeotroped with toluene, and then slurry-washed withn-hexane/ethyl acetate (1/1, 440 ml) to give the titled compound (30.1g).

¹H-NMR (DMSO-D₆) 6:10.24 (1H, br s), 9.64 (2H, br s), 8.90 (1H, br s),7.58-7.53 (2H, m), 7.47-7.41 (3H, m), 4.21-4.10 (2H, m), 4.02-3.94 (1H,m), 3.46-3.37 (1H, m), 3.20-3.10 (1H, m), 2.99-2.85 (2H, m), 2.69-2.54(2H, m), 1.10 (3H, d, J=7.2 Hz).

(10) Optically-Active Compound of6-benzyl-3-methyl-1,6-diazaspiro[3.4]octan-5-one

To a solution of an optically-active compound of2-(2-benzylaminoethyl)-3-methylazetidine-2-dicarboxylic acid 2hydrochloride (29.1 g) and N,N-diisopropylethylamine (65 ml) inchloroform (290 ml) was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (41.3 g) at room temperature, and the mixture wasstirred for 4 hours. To the reaction mixture were added saturatedaqueous sodium bicarbonate solution (200 ml) and water (100 ml), and themixture was extracted with chloroform (200 ml). The organic layer waswashed with saturated aqueous sodium chloride solution, dried overanhydrous magnesium sulfate and concentrated under reduced pressure. Theresulting residue was purified by silica gel column chromatography(eluent: chloroform/methanol=20/1 to 10/1) to give the titled compound(21.3 g).

¹H-NMR (DMSO-D₆) δ: 7.38-7.31 (2H, m), 7.30-7.22 (3H, m), 4.52 (1H, d,J=14.8 Hz), 4.29 (1H, d, J=14.8 Hz), 3.35-3.27 (2H, m), 3.22-3.17 (1H,m), 3.05 (2H, dd, J=9.5, 4.0 Hz), 2.77-2.66 (1H, m), 2.16-2.10 (1H, m),1.96-1.87 (1H, m), 0.94 (3H, d, J=7.1 Hz).

(11) Optically-Active Compound of6-benzyl-3-methyl-1,6-diazaspiro[3.4]octane-1-carboxylic acid tert-butylester

To a suspension of lithium aluminum hydride (6.8 g) in tetrahydrofuran(300 ml) was slowly added dropwise concentrated sulfuric acid (4.8 ml)under ice-cooling, and the mixture was stirred for 30 minutes. To themixture was added dropwise a solution of an optically-active compound of6-benzyl-3-methyl-1,6-diazaspiro[3.4]octan-5-one (21.3 g) intetrahydrofuran (100 ml), and the mixture was stirred at the sametemperature for additional 45 minutes. To the reaction mixture weresequentially added water (7.0 ml), 4M aqueous sodium hydroxide solution(7.0 ml) and water (14.0 ml), and the mixture was directly stirred for30 minutes. To the mixture were added anhydrous magnesium sulfate andethyl acetate (100 ml), and the mixture was stirred, and then filteredthrough Celite. To the filtrate was added di-tert-butyl dicarbonate(23.4 g) at room temperature, and the mixture was stirred for 3 hours.The mixture was concentrated under reduced pressure until reduced byhalf and washed with saturated aqueous ammonium chloride solution (200ml×2) twice. To the separated organic layer was added n-hexane (200 ml),and the mixture was extracted with 10% aqueous citric acid solution 5times. The separated aqueous layer was basified by 4M aqueous sodiumhydroxide solution and extracted with chloroform. The organic layer waswashed with saturated aqueous sodium chloride solution (200 ml), driedover anhydrous magnesium sulfate and concentrated under reducedpressure. The resulting residue was purified by silica gel columnchromatography (eluent: chloroform/methanol=40/1 to 20/1) to give thetitled compound (15.6 g).

¹H-NMR (DMSO-D₆) δ: 7.34-7.27 (4H, m), 7.26-7.21 (1H, m), 3.84-3.69 (1H,m), 3.62-3.47 (2H, m), 3.19-3.05 (1H, m), 3.02-2.92 (1H, m), 2.76-2.69(1H, m), 2.47-2.24 (4H, m), 1.95-1.77 (1H, m), 1.36 (9H, s), 1.03 (3H,d, J=7.0 Hz).

(12) Optically-Active Compound of3-methyl-1,6-diazaspiro[3.4]octane-1-carboxylic acid tert-butyl ester

To a solution of an optically-active compound of6-benzyl-3-methyl-1,6-diazaspiro[3.4]octane-1-carboxylic acid tert-butylester (10.0 g) in tetrahydrofuran/methanol (50 ml/50 ml) was added 20%palladium hydroxide carbon (2.0 g), and the mixture was hydrogenatedunder 4 atmospheres for 24 hours. The mixture was filtered throughCelite, and the filtrate was concentrated under reduced pressure to givethe titled compound (7.3 g).

¹H-NMR (DMSO-D₆) δ: 3.88-3.71 (1H, m), 3.44-3.06 (2H, m), 3.02-2.64 (4H,m), 2.55-2.38 (1H, m), 2.31-2.15 (1H, m), 1.81-1.72 (1H, m), 1.37 (9H,s), 1.07 (3H, d, J=7.0 Hz).

(13) Optically-Active Compound of3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.4]octane-1-carboxylicacid tert-butyl ester

An optically-active compound of3-methyl-1,6-diazaspiro[3.4]octane-1-carboxylic acid tert-butyl ester(6.9 g) was mixed with 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (4.3 g),potassium carbonate (7.7 g) and water (65 ml), and stirred for 4 hourswith refluxing. The mixture was cooled to room temperature, and theretowas added water (60 ml). The mixture was extracted withchloroform/methanol (10/1, 120 ml). The organic layer was sequentiallywashed with water, saturated aqueous ammonium chloride solution andsaturated aqueous sodium chloride solution, and dried over anhydroussodium sulfate. To the mixture was added silica gel (4 g), and themixture was stirred for 10 minutes, filtered through Celite andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (eluent: chloroform/ethylacetate=1/1, followed by chloroform/methanol=50/1 to 20/1) to give thetitled compound (10.0 g).

¹H-NMR (DMSO-D₆) δ: 11.59 (1H, br s), 8.09 (1H, s), 7.12-7.09 (1H, m),6.64-6.59 (1H, m), 4.09-3.66 (5H, m), 3.39-3.21 (1H, m), 2.64-2.44 (2H,m), 2.27-2.06 (1H, m), 1.36 (3H, s), 1.21 (6H, s), 1.11 (3H, d, J=6.5Hz).

(14) Optically-Active Compound of4-(3-methyl-1,6-diazaspiro[3.4]oct-6-yl)-7H-pyrrolo[2,3-d]pyrimidine 2hydrochloride

An optically-active compound of3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.4]octane-1-carboxylicacid tert-butyl ester (9.5 g) was mixed with 4M hydrochloricacid-1,4-dioxane (50 ml), chloroform (50 ml) and methanol (100 ml), andthe mixture was stirred at 60° C. for 30 minutes. The mixture wasconcentrated under reduced pressure, and azeotroped with toluene to givethe titled compound (9.3 g).

¹H-NMR (DMSO-D₆) δ: 12.91 (1H, br s), 9.97-9.64 (2H, m), 8.45-8.35 (1H,m), 7.58-7.47 (1H, m), 7.04-6.92 (1H, m), 4.99-4.65 (1H, m), 4.32-3.21(7H, m), 3.04-2.90 (1H, m), 2.46-2.31 (1H, m), 1.27 (3H, d, J=6.0 Hz).

(15) Optically-Active Compound of3-[3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.4]oct-1-yl]-3-oxopropionitrile

An optically-active compound of4-(3-methyl-1,6-diazaspiro[3.4]oct-6-yl)-7H-pyrrolo[2,3-d]pyrimidine 2hydrochloride (8.8 g) was mixed with 1-cyanoacetyl-3,5-dimethylpyrazole(6.8 g), N,N-diisopropylethylamine (20 ml) and 1,4-dioxane (100 ml), andthe mixture was stirred at 100° C. for 1 hour. The mixture was cooled toroom temperature, and thereto was added saturated aqueous sodiumbicarbonate solution. The mixture was extracted with chloroform/methanol(10/1). The separated organic layer was washed with saturated aqueoussodium chloride solution, dried over anhydrous magnesium sulfate andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (eluent: chloroform/methanol=30/1 to9/1), and concentrated under reduced pressure. The resulting residue wasslurry-washed with n-heptane/ethanol (2/1, 90 ml) to give a solid (7.3g). The solid was slurry-washed with n-heptane/ethanol (5/1, 90 ml)again to give Crystal 1 of the titled compound (6.1 g).

¹H-NMR (DMSO-D₆) δ: 11.60 (1H, br s), 8.08 (1H, s), 7.11 (1H, dd, J=3.5,2.4 Hz), 6.58 (1H, dd, J=3.4, 1.9 Hz), 4.18-4.14 (1H, m), 4.09-3.93 (3H,m), 3.84-3.73 (1H, m), 3.71 (1H, d, J=19.0 Hz), 3.66 (1H, d, J=18.7 Hz),3.58 (1H, dd, J=8.2, 6.0 Hz), 2.70-2.58 (2H, m), 2.24-2.12 (1H, m), 1.12(3H, d, J=7.1 Hz).

[α]D=+47.09° (25° C., c=0.55, methanol)

To the resulting Crystal 1 (2.6 g) was added 1-butanol (39 ml), and themixture was heated at 100° C. and stirred. After the completedissolution of solids, the solution was cooled to room temperature by10° C. per 30 minutes, and further stirred at room temperatureovernight. The generated crystals were filtered, washed with 1-butanol(6.2 ml) and dried under reduced pressure to give Crystal 2 of thetitled compound (2.1 g).

[Preparation 7]: Synthesis of Compound 7

(1) Optically-Active Compound of3-trichloromethyltetrahydropyrrolo[1,2-c]oxazol-1-one

To a solution of D-proline (100.0 g) in acetonitrile (400 ml) was addeddropwise chloral (169 ml) at room temperature, and the mixture wasstirred for 18 hours. The mixture was filtered, and the filtrate wasconcentrated under reduced pressure. To the resulting residue was addedwater, and the mixture was extracted with chloroform. The separatedorganic layer was dried over anhydrous magnesium sulfate, andconcentrated under reduced pressure. The resulting residue wasslurry-washed with n-hexane/ethyl acetate (20/1, 900 ml) to give thetitled compound (159.7 g).

¹H-NMR (DMSO-D₆) δ: 5.83 (1H, s), 4.10 (1H, dd, J=9.0, 4.2 Hz),3.34-3.27 (1H, m), 3.20-3.13 (1H, m), 2.19-2.07 (1H, m), 1.98-1.90 (1H,m), 1.83-1.73 (1H, m), 1.67-1.55 (1H, m).

(2) Optically-Active Compound of7a-allyl-3-trichloromethyltetrahydropyrrolo[1,2-c]oxazol-1-one

To a solution of diisopropylamine (60 ml) in tetrahydrofuran (160 ml)was added dropwise n-butyllithium (2.64M hexane solution, 161 ml) over20 minutes under ice-cooling, and the mixture was directly stirred for40 minutes. To the mixture cooled to −68° C. was added dropwise asolution of 3-trichloromethyltetrahydropyrrolo[1,2-c]oxazol-1-one (80.0g) in tetrahydrofuran (640 ml) over 30 minutes, and the mixture wasdirectly stirred for 20 minutes. To the mixture was added allyl bromide(57 ml), and the mixture was directly stirred for 1 hour. To thereaction mixture was added saturated aqueous ammonium chloride solution(1000 ml), and the mixture was extracted with ethyl acetate (800 ml).The separated organic layer was washed with water (800 ml) and saturatedaqueous sodium chloride solution (500 ml). The separated aqueous layerwas extracted with ethyl acetate (400 ml, 500 ml) twice. The combinedorganic layer was dried over anhydrous magnesium sulfate, andconcentrated under reduced pressure to give the titled compound (86.1g).

¹H-NMR (CDCl₃) δ: 5.96-5.84 (1H, m), 5.22-5.21 (1H, m), 5.20-5.16 (1H,m), 5.00-4.98 (1H, m), 3.26-3.15 (2H, m), 2.67-2.53 (2H, m), 2.19-2.11(1H, m), 2.08-1.98 (1H, m), 1.95-1.85 (1H, m), 1.72-1.61 (1H, m).

(3) Optically-Active Compound of 2-allylpyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester

To a solution of7a-allyl-3-trichloromethyltetrahydropyrrolo[1,2-c]oxazol-1-one (86.1 g)in methanol (430 ml) cooled to 0° C. was added dropwise concentratedsulfuric acid (43 ml), and then the mixture was stirred for 13.5 hourswith refluxing. The mixture was cooled to room temperature andconcentrated under reduced pressure. Then, to the resulting residue wasadded ethyl acetate (500 ml), and the mixture was extracted with water(500 ml). The separated organic layer was extracted with water (300 ml)again. The combined aqueous layer was neutralized by 4M aqueous sodiumhydroxide solution to pH 7.5. To the mixture was added sodiumbicarbonate (55 g), followed by a solution of di-tert-butyl dicarbonate(85.7 g) in tetrahydrofuran (430 ml) at room temperature, and themixture was stirred overnight. To the mixture was added ethyl acetate(800 ml), and the mixture was extracted. The separated organic layer wassequentially washed with water (1000 ml) and saturated aqueous sodiumchloride solution (500 ml). The separated aqueous layer was extractedwith ethyl acetate (400 ml) again. The combined organic layer was driedover anhydrous sodium sulfate, and concentrated under reduced pressure.The resulting residue was purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=30/1 to 10/1) to give the titledcompound (61.2 g).

¹H-NMR (CDCl₃) δ: 5.82-5.70 (1.0H, m), 5.16-5.14 (1.0H, m), 5.13-5.09(1.0H, m), 3.73-3.66 (0.7H, m), 3.72 (3.0H, s), 3.63-3.56 (0.3H, m),3.42-3.31 (1.0H, m), 3.14-3.07 (0.3H, m), 2.96-2.89 (0.7H, m), 2.65-2.57(1.0H, m), 2.17-1.98 (2.0H, m), 1.94-1.75 (2.0H, m), 1.46 (3.0H, s),1.43 (6.0H, s).

(4) Optically-Active Compound of2-(3-hydroxypropyl)pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester2-methyl ester

To a solution of 2-allylpyrrolidine-1,2-dicarboxylic acid 1-tert-butylester 2-methyl ester (25.0 g) in tetrahydrofuran (125 ml) cooled to 0°C. was added dropwise borane-tetrahydrofuran complex (1.0Mtetrahydrofuran solution, 105 ml) over 30 minutes, and the mixture wasstirred at the same temperature for 2.5 hours. To the reaction mixturewas added dropwise additional borane-tetrahydrofuran complex (1.0Mtetrahydrofuran solution, 11 ml), and the mixture was stirred at thesame temperature for 50 minutes. To the reaction mixture was addeddropwise water (180 ml) over 30 minutes and added sodium peroxoborate 1hydrate (12.0 g) in small batches. The mixture was warmed to roomtemperature and stirred overnight, and then thereto was added water (500ml). The mixture was extracted with ethyl acetate (600 ml). Theseparated organic layer was washed with 20% aqueous sodium thiosulfatesolution (600 ml) and saturated aqueous sodium chloride solution. Theseparated aqueous layer was extracted with ethyl acetate (300 ml, 200ml) twice. The combined organic layer was dried over anhydrous sodiumsulfate, and concentrated under reduced pressure. The resulting residuewas purified by silica gel column chromatography (eluent: n-hexane/ethylacetate=2/1 to 1/1) to give the titled compound (17.4 g).

¹H-NMR (CDCl₃) δ: 3.77-3.59 (2H, m), 3.71 (3H, s), 3.47-3.36 (1H, m),2.37-1.76 (7H, m), 1.68-1.50 (3H, m), 1.45 (4H, s), 1.41 (5H, s).

(5) Optically-Active Compound of2-(3-oxopropyl)pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester2-methyl ester

To a suspension of 2-(3-hydroxypropyl)pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester 2-methyl ester (17.4 g) and sodium bicarbonate (12.7g) in chloroform (175 ml) cooled to 0° C. was added Dess-Martinperiodinane (28.3 g). The reaction mixture was warmed to roomtemperature and stirred for 1.5 hours. To the reaction mixture was addedadditional Dess-Martin periodinane (1.0 g), and the mixture was stirredfor 50 minutes. To the reaction mixture cooled to 0° C. was addedadditional Dess-Martin periodinane (15 g), and the mixture was stirredfor 2 hours. To the mixture were added 20% sodium thiosulfate (250 ml)and saturated aqueous sodium bicarbonate solution (250 ml), and themixture was stirred at room temperature for 20 minutes. The mixture wasextracted with chloroform (200 ml×2) twice, and the combined organiclayer was dried over anhydrous sodium sulfate and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (eluent: n-hexane/ethyl acetate=4/1 to 3/2) togive the titled compound (5.68 g).

¹H-NMR (CDCl₃) δ: 9.78-9.76 (0.5H, m), 9.70-9.68 (0.5H, m), 3.76-3.69(3.5H, m), 3.61-3.52 (0.5H, m), 3.44-3.32 (1.0H, m), 2.68-2.40 (3.0H,m), 2.29-2.06 (2.0H, m), 2.03-1.76 (3.0H, m), 1.44 (4.0H, s), 1.41(5.0H, s).

(6) Optically-Active Compound of2-(3-benzylaminopropyl)pyrrolidine-1,2-dicarboxylic acid 1-tert-butylester 2-methyl ester

To a solution of 2-(3-oxopropyl)pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester 2-methyl ester (5.68 g) in tetrahydrofuran (57 ml)was added benzylamine (6.53 ml) at room temperature, and the mixture wasstirred for 1 hour. The mixture was cooled to 0° C., and thereto wasadded sodium triacetoxyborohydride (5.07 g). The mixture was stirred atroom temperature for 12.5 hours. To the reaction mixture were addedwater (12 ml) and saturated aqueous ammonium chloride solution (200 ml),and the mixture was extracted with n-hexane/ethyl acetate (1/2, 210m1×2, 120 m1×2) 4 times. To the combined organic layer was addedn-hexane (150 ml), and the mixture was extracted with 10% aqueouspotassium bisulfate solution (180 ml×3) 3 times. The combined aqueouslayer was neutralized by 4M aqueous sodium hydroxide solution to pH 7 to8, and thereto was added saturated aqueous sodium bicarbonate solution(100 ml). The mixture was extracted with ethyl acetate (200 ml). Theseparated aqueous layer was extracted with ethyl acetate (200 ml×2)twice, and the combined organic layer was washed with saturated aqueoussodium chloride solution (200 ml). The organic layer was dried overanhydrous sodium sulfate and concentrated under reduced pressure to givethe titled compound (7.69 g).

¹H-NMR (CDCl₃) δ: 7.27-7.35 (5H, m), 3.80 (2H, s), 3.65-3.75 (0.7H, m),3.69 (3H, s), 3.55-3.65 (0.5H, m), 3.31-3.45 (1H, m), 2.60-2.71 (2H, m),2.25-2.35 (0.2H, m), 2.10-2.21 (0.5H, m), 2.04 (1.0H, s), 1.97-2.11(2.4H, m), 1.74-1.93 (3.0H, m), 1.45-1.65 (1.7H, m), 1.44 (3.5H, s),1.38 (5.5H, s)

(7) Optically-Active Compound of2-(3-benzylaminopropyl)pyrrolidine-2-carboxylic acid methyl ester

To a solution of 2-(3-benzylaminopropyl)pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (7.69 g) in chloroform (38 ml)cooled to 0° C. was added 4M hydrochloric acid-ethyl acetate (18 ml),and the mixture was stirred at room temperature for 3 hours. To themixture was added additional 4M hydrochloric acidethyl acetate (20 ml),and the mixture was stirred at room temperature for 50 minutes. Thereaction mixture was concentrated under reduced pressure, and thenthereto was added saturated aqueous sodium bicarbonate solution (150ml). The mixture was extracted with chloroform (100 m1×2, 75 m1×2) 4times. The combined organic layer was dried over anhydrous sodiumsulfate and concentrated under reduced pressure to give the titledcompound (7.49 g).

¹H-NMR (CDCl₃) δ: 7.35-7.22 (5H, m), 3.77 (2H, s), 3.71 (3H, s),3.02-2.92 (2H, m), 2.65-2.58 (2H, m), 2.21-2.13 (1H, m), 1.92-1.50 (8H,m), 1.43-1.31 (1H, m).

(8) Optically-Active Compound of 7-benzyl-1,7-diazaspiro[4.5]decan-6-one

A solution of 2-(3-benzylaminopropyl)pyrrolidine-2-carboxylic acidmethyl ester (133 mg) in xylene (1.5 ml) was stirred at 130° C.overnight. The mixture was cooled to room temperature, and purified bysilica gel column chromatography (eluent: chloroform/methanol=10/1) togive the titled compound (90 mg).

¹H-NMR (CDCl₃) δ: 7.35-7.21 (5H, m), 4.68 (1H, d, J=14.6 Hz), 4.48 (1H,d, J=14.6 Hz), 3.31-3.18 (3H, m), 2.90-2.83 (1H, m), 2.12-2.04 (1H, m),1.99-1.80 (7H, m), 1.78-1.70 (1H, m).

(9) Optically-Active Compound of7-benzyl-6-oxo-1,7-diazaspiro[4.5]decane-1-carboxylic acid tert-butylester

To a suspension of lithium aluminum hydride (45 mg) in tetrahydrofuran(1.5 ml) cooled to 0° C. was added concentrated sulfuric acid (31 μl),and the mixture was stirred for 50 minutes. To the mixture was addeddropwise a solution of 7-benzyl-1,7-diazaspiro[4.5]decan-6-one (90 mg)in tetrahydrofuran (0.5 ml) at 0° C., and the mixture was stirred at thesame temperature for 40 minutes. To the reaction mixture weresequentially added water (45 μl), 4M aqueous sodium hydroxide solution(45 μl) and water (45 μl), and the mixture was stirred for 30 minutes.To the mixture were added ethyl acetate and anhydrous magnesium sulfate,and the mixture was filtered through Celite. To the filtrate was addeddi-tert-butyl dicarbonate (112 mg) at room temperature, and the mixturewas stirred for 100 minutes. The reaction mixture was concentrated underreduced pressure, and purified by silica gel column chromatography(eluent: n-hexane/ethyl acetate=10/1 to 5/1) to give the titled compound(76 mg).

¹H-NMR (DMSO-D₆) δ: 7.36-7.28 (4H, m), 7.25-7.20 (1H, m), 3.72-3.67 (2H,m), 3.62 (2H, s), 3.58 (1H, s), 3.56-3.45 (1H, m), 3.31-3.22 (1H, m),2.55-2.47 (1H, m), 2.17-1.90 (3H, m), 1.83-1.72 (3H, m), 1.54-1.41 (1H,m), 1.37 (3H, s), 1.33-1.23 (1H, m), 1.29 (6H, s).

(10) Optically-Active Compound of 1,7-diazaspiro[4.5]decane-1-carboxylicacid tert-butyl ester

To a solution of 7-benzyl-6-oxo-1,7-diazaspiro[4.5]decane-1-carboxylicacid tert-butyl ester (76 mg) in tetrahydrofuran/methanol (1 ml/1 ml)was added 20% palladium hydroxide carbon (15 mg), and the mixture washydrogenated under 4 atmospheres. The mixture was filtered throughCelite under nitrogen, and the filtrate was concentrated under reducedpressure to give the titled compound (51 mg).

¹H-NMR (CDCl₃) δ: 3.57-3.31 (3H, m), 3.00-2.87 (1H, m), 2.78-2.39 (3H,m), 2.17-2.07 (1H, m), 1.93-1.78 (1H, m), 1.76-1.62 (3H, m), 1.53-1.42(12H, m).

(11) Optically-Active Compound of7-(7H-pyrrolo[2.3-d]pyrimidin-4-yl)-1,7-diazaspiro[4.5]decane-1-carboxylicacid tert-butyl ester

1,7-Diazaspiro[4.5]decane-1-carboxylic acid tert-butyl ester (51 mg) wasmixed with 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (36 mg), potassiumcarbonate (59 mg) and water (1 ml), and the mixture was stirred for 1.5hours with refluxing. The mixture was cooled to room temperature, andthereto was added water. The mixture was extracted with chloroform. Theorganic layer was washed with saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfate and concentrated underreduced pressure. The resulting residue was purified by silica gelcolumn chromatography (eluent: n-hexane/ethyl acetate=1/1, followed bychloroform/methanol=20/1) to give the titled compound (60 mg).

¹H-NMR (CDCl₃) δ: 10.58-10.90 (1H, m), 8.30 (1H, br s), 7.07 (1H, br s),6.48-6.56 (1H, m), 4.66-4.87 (1H, m), 4.52-4.62 (1H, m), 3.82-3.95(0.6H, m), 3.50-3.75 (1.4H, m), 3.31-3.50 (1H, m), 3.04-3.20 (0.6H, m),2.85-3.04 (1H, m), 2.58-2.74 (0.4H, m), 1.98-2.14 (1H, m), 1.40-1.90(6H, m), 1.54 (3.6H, s), 1.48 (5.4H, s)

(12) Optically-Active Compound of4-(1,7-diazaspiro[4.5]dec-7-yl)-7H-pyrrolo[2.3-d]pyrimidine

To a solution of7-(7H-pyrrolo[2.3-d]pyrimidin-4-yl)-1,7-diazaspiro[4.5]decane-1-carboxylicacid tert-butyl ester (60 mg) in chloroform (1.0 ml) were added 4Mhydrochloric acid-ethyl acetate (1.5 ml) and 2M hydrochloricacid-methanol (0.5 ml), and the mixture was stirred at room temperaturefor 2.5 hours. The reaction mixture was concentrated under reducedpressure, and then azeotroped with toluene. The residue was neutralizedby the addition of 4M aqueous sodium hydroxide solution, and thereto wasadded saturated aqueous sodium chloride solution. The mixture wasextracted with chloroform. The separated organic solvent wasconcentrated under reduced pressure to give the titled compound (30 mg).

¹H-NMR (CDCl₃) δ: 9.26 (1H, br s), 8.28 (1H, s), 7.04 (1H, d, J=3.7 Hz),6.52 (1H, d, J=3.7 Hz), 3.92-3.86 (2H, m), 3.81 (1H, d, J=13.0 Hz), 3.72(1H, d, J=13.0 Hz), 3.10-3.00 (2H, m), 1.94-1.80 (3H, m), 1.78-1.70 (5H,m), 1.55-1.47 (1H, m).

(13) Optically-Active Compound of3-oxo-3-[7-(7H-pyrrolo[2.3-d]pyrimidin-4-yl)-1,7-diazaspiro[4.5]dec-1-yl)propionitrile

4-(1,7-Diazaspiro[4.5]dec-7-yl)-7H-pyrrolo[2.3-d]pyrimidine (30 mg) wasmixed with 1-cyanoacetyl-3,5-dimethylpyrazole (38 mg),N,N-diisopropylethylamine (42 μl) and 1,4-dioxane (1 ml), and themixture was stirred at 110° C. for 75 minutes. The mixture was cooled toroom temperature, and then thereto was added saturated aqueous sodiumbicarbonate solution. The mixture was extracted with ethyl acetate. Theorganic layer was dried over anhydrous magnesium sulfate, andconcentrated under reduced pressure. The resulting residue was purifiedby silica gel column chromatography (eluent: chloroform/ethylacetate=1/2, followed by chloroform/methanol=25/1 to 20/1) to give thetitled compound (32 mg).

¹H-NMR (DMSO-D₆) δ: 11.69 (1H, br s), 8.09 (1H, s), 7.19-7.15 (1H, m),6.59-6.55 (1H, m), 4.74-4.66 (1H, m), 4.55-4.47 (1H, m), 3.95 (2H, s),3.86-3.78 (1H, m), 3.57-3.48 (1H, m), 3.45-3.37 (1H, m), 2.99-2.82 (2H,m), 1.89-1.68 (4H, m), 1.66-1.56 (2H, m), 1.55-1.47 (1H, m).

[α]D=+185.580 (25° C., c=1.04, methanol)

Preparation 8

An optical active compound of3-[3-methyl-6-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1,6-diazaspiro[3.4]oct-1-yl]-3-oxopropionitrile(Compound 6) was treated according to a conventional method to give 1hydrate thereof.

¹H-NMR (DMSO-D₆) δ: 11.60 (1H, br s), 8.08 (1H, s), 7.11 (1H, s), 6.58(1H, s), 4.16 (1H, dd, J=8.2, 8.2 Hz), 4.11-3.61 (6H, m), 3.57 (1H, dd,J=7.65, 6.26 Hz), 2.70-2.57 (2H, m), 2.24-2.10 (1H, m), 1.11 (3H, d,J=6.9 Hz).

The following Tables 1 to 3 show structural formulae and ¹H-NMR spectraldata of Compounds 1 to 95 prepared according to the above Preparations.¹H-NMR spectra are measured in CDCl₃ or DMSO-d₆ by usingtetramethylsilane as an internal standard, and all δ values are shown inppm. Unless otherwise specified, a 400 MHz NMR spectroscopy was used formeasurement.

Symbols in Table have the following meanings.

s: singletd: doublett: tripletq: quartetdd: double doubletddd: double double doubletbrs: broad singletm: multipletJ: coupling constant

TABLE 1-1 No. Structural Formula Comments NMR Data  1

Compound 1 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.71 (1H,br s), 8.12 (1H, s), 7.20 (1H, dd, J = 3.5, 2.4 Hz), 6.65 (1H, dd, J =3.6, 1.9 Hz), 4.93-4.88 (1H, m), 4.64-4.58 (1H, m), 4.24-4.18 (1H, m),3.67 (2H, s), 3.61 (1H, d, J = 12.8 Hz), 3.46-3.41 (1H, m), 3.03-2.95(1H, m), 2.42-2.35 (1H, m), 2.34-2.25 (1H, m), 2.15-2.08 (1H, m),1.83-1.77 (1H, m), 1.57-1.43 (1H, m), 1.01 (3H, d, J = 7.1 Hz).  2

Compound 2 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.70 (1H,br s), 8.14 (1H, s), 7.19 (1H, dd, J = 3.5, 2.4 Hz), 6.64 (1H, dd, J =3.6, 1.9 Hz), 5.03-4.96 (1H, m), 4.66-4.59 (1H, m), 4.11-4.06 (1H, m),3.70 (1H, d, J = 18.7 Hz), 3.65 (1H, d, J = 18.7 Hz), 3.60- 3.55 (1H,m), 3.45 (1H, d, J = 13.0 Hz), 2.97-2.89 (1H, m), 2.46-2.41 (1H, m),2.40-2.34 (1H, m), 2.00-1.94 (1H, m), 1.88-1.80 (1H, m), 1.64-1.51 (1H,m), 0.91 (3H, d, J = 7.1 Hz).  3

Compound 3 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.73 (1H,br s), 8.13 (1H, s), 7.22-7.18 (1H, m), 6.67-6.63 (1H, m), 4.96 (1H, d,J = 12.6 Hz), 4.68-4.60 (1H, m), 4.11-4.03 (1H, m), 4.01-3.93 (1H, m),3.71 (2H, s), 3.53 (1H, d, J = 12.6 Hz), 2.98-2.88 (1H, m), 2.40-2.30(1H, m), 2.03-1.90 (3H, m), 1.83-1.75 (1H, m), 1.59-1.45 (1H, m).  4Compound 4 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.68 (1H,br s), 8.11 (1H, s), 7.17-7.14 (1H, m), 6.58 (1H, dd, J = 3.3, 1.8 Hz),4.66-4.58 (2H, m), 4.24-4.13 (2H, m), 4.12-4.02 (1H, m), 3.89-3.78 (3H,m), 2.68-2.58 (1H, m), 2.56-2.45 (1H, m).  5

Compound 5 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.74 (1H,br s), 8.15 (1H, s), 7.22-7.19 (1H, m), 6.65-6.62 (1H, m), 5.13-5.07(1H, m), 4.69-4.46 (3H, m), 3.83 (1H, d, J = 18.8 Hz), 3.77 (1H, d, J =19.2 Hz), 3.61- 3.55 (1H, m), 2.99-2.91 (1H, m), 2.39- 2.29 (1H, m),2.26-2.19 (1H, m), 1.99- 1.91 (1H, m), 1.63-1.48 (1H, m).  6

Compound 6 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.60 (1H,br s), 8.08 (1H, s), 7.11 (1H, dd, J = 3.5, 2.4 Hz), 6.58 (1H, dd, J =3.4, 1.9 Hz), 4.18-4.14 (1H, m), 4.09-3.93 (3H, m), 3.84-3.73 (1H, m),3.71 (1H, d, J = 19.0 Hz), 3.66 (1H, d, J = 18.7 Hz), 3.58 (1H, dd, J =8.2, 6.0 Hz), 2.70-2.58 (2H, m), 2.24-2.12 (1H, m), 1.12 (3H, d, J = 7.1Hz).  7

Compound 7 (Optically-active substance) 1H-NMR (DMSO-D6) δ: 11.69 (1H,br s), 8.09 (1H, s), 7.19-7.15 (1H, m), 6.59-6.55 (1H, m), 4.74-4.66(1H, m), 4.55-4.47 (1H, m), 3.95 (2H, s), 3.86- 3.78 (1H, m), 3.57-3.48(1H, m), 3.45- 3.37 (1H, m), 2.99-2.82 (2H, m), 1.89- 1.68 (4H, m),1.66-1.56 (2H, m), 1.55- 1.47 (1H, m).  8

Racemate 1H-NMR (DMSO-D6) δ: 11.66-11.52 (1.0H, m), 8.28 (0.1H, s), 8.10(0.3H, s), 8.08-8.05 (0.6H, m), 7.15-7.06 (1.0H, m), 6.63-6.51 (1.0H,m), 4.49-4.16 (0.5H, m), 4.06-3.74 (2.0H, m), 3.66- 2.72 (9.5H, m),2.44-2.32 (0.5H, m), 2.15-1.65 (6.5H, m).  9

Enantiomer of Compound 4 1H-NMR (DMSO-D6) δ: 11.68 (1H, br s), 8.11 (1H,s), 7.17-7.15 (1H, m), 6.58 (1H, dd, J = 3.2, 1.7 Hz), 4.65-4.59 (2H,m), 4.22-4.15 (2H, m), 4.13-4.01 (2H, m), 3.86 (1H, d, J = 19.0 Hz),3.80 (1H, d, J = 19.2 Hz), 2.69-2.58 (1H, m), 2.56-2.45 (1H, m). 10

Enantiomer of Compound 6 1H-NMR (DMSO-D6) δ: 11.60 (1H, br s), 8.08 (1H,s), 7.11 (1H, dd, J = 3.2, 2.5 Hz), 6.58 (1H, dd, J = 3.4, 1.9 Hz),4.18-4.14 (1H, m), 4.09-3.93 (3H, m), 3.86-3.74 (1H, m), 3.71 (1H, d, J= 18.7 Hz), 3.66 (1H, d, J = 19.0 Hz), 3.58 (1H, dd, J = 8.2, 6.0 Hz),2.69-2.58 (2H, m), 2.24-2.12 (1H, m), 1.12 (3H, d, J = 7.1 Hz). 11

Enantiomer of Compound 2 1H-NMR (DMSO-D6) δ: 11.70 (1H, br s), 8.14 (1H,s), 7.19 (1H, dd, J = 3.5, 2.4 Hz), 6.64 (1H, dd, J = 3.7, 2.0 Hz),5.02-4.97 (1H, m), 4.66-4.59 (1H, m), 4.11-4.06 (1H, m), 3.70 (1H, d, J= 18.7 Hz), 3.65 (1H, d, J = 19.0 Hz), 3.60- 3.56 (1H, m), 3.45 (1H, d,J = 13.0 Hz), 2.98-2.89 (1H, m), 2.46-2.41 (1H, m), 2.39-2.34 (1H, m),2.00-1.94 (1H, m), 1.87-1.81 (1H, m), 1.64-1.51 (1H, m), 0.91 (3H, d, J= 7.1 Hz). 12

Enantiomer of Compound 1 1H-NMR (DMSO-D6) δ: 11.71 (1H, br s), 8.13 (1H,s), 7.20 (1H, dd, J = 3.5, 2.4 Hz), 6.65 (1H, dd, J = 3.5, 1.8 Hz),4.93-4.88 (1H, m), 4.64-4.58 (1H, m), 4.24-4.18 (1H, m), 3.68 (2H, s),3.61 (1H, d, J = 12.8 Hz), 3.45-3.41 (1H, m), 3.04-2.95 (1H, m),2.42-2.35 (1H, m), 2.34-2.25 (1H, m), 2.15-2.08 (1H, m), 1.85-1.76 (1H,m), 1.57-1.43 (1H, m), 1.01 (3H, d, J = 7.1 Hz). 13

Diastereomer of Compound 34 (Optically-active substance) 1H-NMR(DMSO-D6) δ: 11.56 (1H, br s), 8.07 (1H, s), 7.12-7.08 (1H, m),6.50-6.46 (1H, m), 4.84 (1H, q, J = 6.5 Hz), 4.05-3.98 (1H, m), 3.92(2H, s), 3.82-3.74 (1H, m), 3.51-3.44 (2H, m), 2.68-2.59 (1H, m),2.20-2.13 (1H, m), 1.97-1.90 (1H, m), 1.89-1.79 (3H, m), 1.26 (3H, d, J= 6.5 Hz). 14

Diastereomer of Compound 20 (Racemate) 1H-NMR (DMSO-D6) δ: 11.56 (1H, brs), 8.13-8.09 (1H, m), 7.21-7.16 (1H, m), 6.67-6.63 (1H, m), 4.91-4.84(1H, m), 4.69-4.61 (1H, m), 4.51-4.44 (1H, m), 3.77 (1H, d, J = 18.6Hz), 3.69 (1H, d, J = 18.6 Hz), 3.65 (1H, d, J = 15.3 Hz), 2.94-2.85(1H, m), 2.38-2.29 (1H, m), 2.22-2.16 (1H, m), 1.97-1.91 (1H, m),1.79-1.71 (1H, m), 1.58-1.48 (2H, m), 1.34 (3H, d, J = 6.3 Hz). 15

Racemate of Compound 1 1H-NMR (CDCl3) δ: 9.35 (1H, br s), 8.30 (1H, s),7.07 (1H, dd, J = 3.6, 2.3 Hz), 6.53 (1H, dd, J = 3.6, 1.9 Hz),5.09-5.03 (1H, m), 4.65-4.59 (1H, m), 4.35-4.30 (1H, m), 3.81-3.76 (1H,m), 3.60-3.55 (1H, m), 3.21-3.13 (1H, m), 3.21 (2H, s), 2.74-2.65 (1H,m), 2.59- 2.49 (1H, m), 2.23-2.17 (1H, m), 1.93- 1.86 (1H, m), 1.72-1.60(1H, m), 1.13 (3H, d, J = 7.1 Hz). 16

Single diastereomer (Racemate) 1H-NMR (DMSO-D6) δ: 11.73 (1H, br s),8.12 (1H, s), 7.22-7.19 (1H, m), 6.65-6.62 (1H, m), 4.99-4.92 (1H, m),4.64-4.57 (1H, m), 4.09-4.01 (1H, m), 4.00-3.92 (1H, m), 3.69 (2H, s),3.49- 3.44 (1H, m), 2.60-2.52 (1H, m), 2.08- 1.90 (4H, m), 1.75-1.65(1H, m), 0.98 (3H, d, J = 6.5 Hz). 17

Racemate 1H-NMR (DMSO-D6) δ: 11.61 (1H, br s), 8.08 (1H, s), 7.12 (1H,dd, J = 3.3, 2.4 Hz), 6.57 (1H, dd, J = 3.5, 1.8 Hz), 4.29-4.20 (1H, m),4.08-3.74 (5H, m), 3.70 (2H, s), 2.69-2.58 (1H, m), 2.43- 2.36 (1H, m),2.35-2.27 (1H, m), 2.23- 2.14 (1H, m). 18

Single diastereomer (Racemate) 1H-NMR (CDCl3) δ: 10.33 (1H, br s), 8.31(1H, s), 7.09 (1H, dd, J = 3.5, 2.0 Hz), 6.53-6.49 (1H, m), 5.08-4.99(1H, m), 4.97-4.88 (1H, m), 4.29-4.21 (1H, m), 4.18-4.11 (1H, m),3.88-3.81 (1H, m), 3.25 (2H, s), 2.88-2.79 (1H, m), 2.44-2.36 (1H, m),2.10-2.02 (1H, m), 1.92-1.79 (2H, m), 1.78-1.71 (1H, m), 1.42 (3H, d, J= 7.1 Hz). 19

Diastereomer of Compound 6 (Racemate) 1H-NMR (DMSO-D6) δ: 11.57 (1H, brs), 8.07 (1H, s), 7.11 (1H, dd, J = 3.6, 2.4 Hz), 6.57 (1H, dd, J = 3.4,1.8 Hz), 4.31-4.22 (1H, m), 4.18-4.13 (1H, m), 4.09-3.94 (1H, m),3.91-3.73 (2H, m), 3.67 (2H, s), 3.59-3.53 (1H, m), 2.80- 2.71 (1H, m),2.45-2.36 (1H, m), 2.30- 2.19 (1H,m), 1.15 (3H, d, J = 6.9 Hz). 20

Diastereomer of Compound 14 (Racemate) 1H-NMR (CDCl3) δ: 9.04 (1H, brs), 8.32 (1H, s), 7.09-7.06 (1H, m), 6.56- 6.53 (1H, m), 5.10-5.05 (1H,m), 4.72- 4.66 (1H, m), 4.53-4.47 (1H, m), 3.78- 3.73 (1H, m), 3.30 (1H,d, J = 17.2 Hz), 3.25 (1H, d, J = 17.6 Hz), 3.12-3.04 (1H, m), 2.77-2.69(1H, m), 2.23-2.17 (1H, m), 1.95-1.85 (3H, m), 1.59 (4H, d, J = 6.3 Hz).21

Racemate of Compound 6 1H-NMR (DMSO-D6) δ: 11.59 (1H, br s), 8.07 (1H,s), 7.11 (1H, dd, J = 3.2, 2.6 Hz), 6.57 (1H, dd, J = 3.4, 1.7 Hz),4.18-4.12 (1H, m), 4.08-3.92 (3H, m), 3.84-3.72 (1H, m), 3.70 (1H, d, J= 18.8 Hz), 3.65 (1H, d, J = 18.8 Hz), 3.59- 3.54 (1H, m), 2.68-2.58(2H, m), 2.22- 2.11 (1H, m), 1.11 (3H, d, J = 7.2 Hz). 22

Enantiomer of Compound 7 1H-NMR (DMSO-D6) δ: 11.69 (1H, br s), 8.09 (1H,s), 7.17 (1H, dd, J = 3.5, 2.6 Hz), 6.57 (1H, dd, J = 3.5, 1.9 Hz),4.73-4.66 (1H, m), 4.55-4.48 (1H, m), 3.95 (2H, s), 3.85-3.78 (1H, m),3.56- 3.49 (1H, m), 3.45-3.37 (1H, m), 2.98- 2.82 (2H, m), 1.89-1.69(4H, m), 1.65- 1.56 (2H, m), 1.55-1.48 (1H, m). 23

Racemate of Compound 2 1H-NMR (DMSO-D6) δ: 11.70 (1H, br s), 8.14 (1H,s), 7.18 (1H, dd, J = 3.7, 2.6 Hz), 6.64 (1H, dd, J = 3.7, 1.9 Hz),5.02-4.96 (1H, m), 4.65-4.59 (1H, m), 4.11-4.05 (1H, m), 3.70 (1H, d, J= 18.8 Hz), 3.65 (1H, d, J = 18.8 Hz), 3.59- 3.54 (1H, m), 3.46-3.42(1H, m), 2.97- 2.88 (1H, m), 2.47-2.33 (2H, m), 2.00- 1.93 (1H, m),1.87-1.79 (1H, m), 1.63- 1.50 (1H, m), 0.90 (3H, d, J = 7.2 Hz). 24

Single diastereomer (Racemate) 1H-NMR (DMSO-D6) δ: 11.59 (1H, br s),8.09 (1H, s), 7.12 (1H, dd, J = 3.2, 2.6 Hz), 6.52 (1H, dd, J = 3.5, 1.9Hz), 4.45 (1H, q, J = 6.4 Hz), 4.08-3.99 (3H, m), 3.81 (1H, d, J = 18.8Hz), 3.81-3.71 (1H, m), 3.75 (1H, d, J = 18.8 Hz), 3.01- 2.92 (1H, m),2.31-2.24 (1H, m), 2.20- 2.12 (1H, m), 2.09-2.01 (1H, m), 1.33 (3H, d, J= 6.3 Hz). 25

Enantiomer of Compound 3 1H-NMR (DMSO-D6) δ: 11.72 (1H, br s), 8.13 (1H,s), 7.20 (1H, dd, J = 3.5, 2.6 Hz), 6.64 (1H, dd, J = 3.6, 1.9 Hz), 4.96(1H, d, J = 12.6 Hz), 4.68-4.59 (1H, m), 4.11-4.04 (1H, m), 4.00-3.93(1H, m), 3.71 (2H, s), 3.53 (1H, d, J = 12.6 Hz), 2.98-2.90 (1H, m),2.39-2.30 (1H, m), 2.01-1.93 (3H, m), 1.82-1.75 (1H, m), 1.59-1.46 (1H,m). 26

Racemate of Compound 4 1H-NMR (DMSO-D6) δ: 11.67 (1H, br s), 8.11 (1H,s), 7.16 (1H, dd, J = 3.3, 2.4 Hz), 6.58 (1H, dd, J = 3.5, 2.0 Hz),4.65-4.57 (2H, m), 4.23-4.14 (2H, m), 4.12-4.01 (1H, m), 3.89-3.78 (1H,m), 3.85 (1H, d, J = 19.2 Hz), 3.79 (1H, d, J = 19.0 Hz), 2.66-2.58 (1H,m), 2.56- 2.46 (1H, m). 27

Enantiomer of Compound 5 1H-NMR (DMSO-D6) δ: 11.74 (1H, br s), 8.15 (1H,s), 7.22-7.19 (1H, m), 6.65-6.62 (1H, m), 5.13-5.07 (1H, m), 4.70-4.46(3H, m), 3.83 (1H, d, J = 18.8 Hz), 3.77 (1H, d, J = 19.0 Hz), 3.61-3.54 (1H, m), 2.99-2.90 (1H, m), 2.39- 2.29 (1H, m), 2.26-2.19 (1H, m),1.99- 1.90 (1H, m), 1.64-1.48 (1H, m). 28

Racemate 1H-NMR (DMSO-D6) δ: 11.70 (1H, br s), 8.15 (1H, s), 7.19 (1H,dd, J = 3.6, 2.5 Hz), 6.65 (1H, dd, J = 3.6, 1.9 Hz), 5.11-5.03 (1H, m),4.60-4.52 (1H, m), 3.78 (1H, d, J = 7.9 Hz), 3.70 (1H, d, J = 18.7 Hz),3.65 (1H, d, J = 18.7 Hz), 3.52 (1H, d, J = 7.9 Hz), 3.50-3.46 (1H, m),3.07-2.99 (1H, m), 2.34-2.25 (1H, m), 2.23-2.18 (1H, m), 1.91-1.83 (1H,m), 1.57-1.44 (1H, m), 1.23 (3H, s), 0.99 (3H, s). 29

Racemate 1H-NMR (CDCl3) δ: 9.43 (1H, br s), 8.28 (1H, s), 7.04 (1H, dd,J = 3.5, 2.1 Hz), 6.52 (1H, dd, J = 3.6, 1.5 Hz), 5.99-5.89 (1H, m),5.33-5.27 (1H, m), 5.23-5.18 (1H, m), 4.75-4.68 (1H, m), 4.62-4.55 (1H,m), 4.14-4.08 (1H, m), 4.05-4.01 (2H, m), 3.81-3.76 (2H, m), 3.68-3.61(1H, m), 3.56-3.48 (1H, m), 3.21-3.11 (2H, m), 2.66-2.54 (2H, m),2.12-2.04 (1H, m), 1.95-1.85 (2H, m), 1.83-1.65 (3H, m), 1.58-1.51 (1H,m). 30

Enantiomer of Compound 36. Configuration of α- carbon of carbonyl is S(derived from reagents). 1H-NMR (CDCl3) δ: 9.54 (1H, br s), 8.29 (1H,s), 7.06 (1H, dd, J = 3.6, 1.7 Hz), 6.54-6.50 (1H, m), 4.71-4.64 (2H,m), 4.30-4.23 (1H, m), 4.06-4.00 (1H, m), 3.74 (1H, br s), 3.57-3.51(2H, m), 3.29-3.21 (1H, m), 3.19-3.10 (1H, m), 2.11-2.04 (1H, m),1.98-1.72 (5H, m), 1.62-1.56 (1H, m), 1.38 (3H, d, J = 6.5 Hz). 31

Racemate 1H-NMR (DMSO-D6) δ: 11.79 (1H, br s), 8.15 (1H, s), 7.23 (1H,dd, J = 3.5, 2.6 Hz), 6.66 (1H, dd, J = 3.5, 1.9 Hz), 4.83-4.76 (1H, m),4.73-4.61 (2H, m), 4.11 (1H, d, J = 19.0 Hz), 4.00 (1H, d, J = 19.2 Hz),3.64-3.50 (2H, m), 3.28- 3.18 (1H, m), 2.35-2.26 (2H, m), 2.11- 2.02(1H, m), 1.93-1.83 (1H, m), 1.79- 1.71 (2H, m). 32

Racemate 1H-NMR (DMSO-D6) δ: 11.73 (1H, s), 8.13 (1H, s), 7.19-7.16 (1H,m), 6.64- 6.60 (1H, m), 4.52-3.96 (4H, m), 4.14 (1H, d, J = 19.0 Hz),4.00 (1H, d, J = 19.2 Hz), 3.61-3.54 (2H, m), 2.46-2.42 (1H, m),2.11-2.01 (1H, m), 1.99-1.91 (1H, m), 1.87-1.76 (1H, m). 33

Diastereomer of Compound 42 (Optically-active substance) 1H-NMR(DMSO-D6) δ: 11.60 (1H, br s), 8.10 (1H, s), 7.13-7.11 (1H, m),6.45-6.43 (1H, m), 4.44-4.31 (2H, m), 4.00 (2H, s), 3.68-3.59 (1H, m),3.52-3.39 (2H, m), 2.87-2.80 (1H, m), 1.99-1.90 (1H, m), 1.83-1.73 (4H,m), 1.36 (3H, d, J = 6.0 Hz). 34

Diastereomer of Compound 13 (Optically-active substance) 1H-NMR(DMSO-D6) δ: 11.59 (1H, br s), 8.08 (1H, s), 7.13-7.10 (1H, m),6.57-6.52 (1H, m), 4.24-4.16 (1H, m), 4.08 (1H, d, J = 19.0 Hz), 4.00(1H, d, J = 19.0 Hz), 3.79-3.67 (2H, m), 3.59-3.53 (1H, m), 3.52-3.44(1H, m), 1.91-1.65 (6H, m), 1.23 (3H, d, J = 6.7 Hz). 35

Diastereomer of Compound 36 (Optically-active substance). Configurationof α- carbon of carbonyl is S (derived from reagents). Higher polarityon TLC (chloroform/methanol = 10/1) than Compound 36. 1H-NMR (CDCl3) δ:9.88 (1H, br s), 8.30 (1H, s), 7.09-7.05 (1H, m), 6.54-6.51 (1H, m),4.79-4.72 (1H, m), 4.63 (1H, d, J = 12.3 Hz), 4.33 (1H, dt, J = 6.6, 6.6Hz), 4.10 (1H, d, J = 12.5 Hz), 3.88 (1H, br s), 3.72- 3.65 (1H, m),3.46-3.38 (1H, m), 3.24-3.16 (1H, m), 3.14-3.06 (1H, m), 2.21-2.14 (1H,m), 2.03-1.52 (6H, m), 1.34 (3H, d, J = 6.5 Hz). 36

Diastereomer of Compound 35, Enantiomer of Compound 30. Configuration ofα- carbon of carbonyl is R (derived from reagents). Lower polarity onTLC (chloroform/methanol = 10/1) than Compound 35. 1H-NMR (CDCl3) δ:10.07 (1H, br s), 8.30 (1H, s), 7.10-7.06 (1H, m), 6.54-6.50 (1H, m),4.73-4.64 (2H, m), 4.27 (1H, dt, J = 6.5, 6.5 Hz), 4.07-4.01 (1H, m),3.77 (1H, br s), 3.60-3.51 (2H, m), 3.30-3.21 (1H, m), 3.19-3.10 (1H,m), 2.12-2.04 (1H, m), 1.98-1.57 (6H, m), 1.38 (3H, d, J = 6.5 Hz). 37

Racemate Compound 5 1H-NMR (DMSO-D6) δ: 11.73 (1H, br s), 8.15 (1H, s),7.21 (1H, dd, J = 3.4, 2.6 Hz), 6.64 (1H, dd, J = 3.6, 2.0 Hz),5.14-5.07 (1H, m), 4.70-4.46 (3H, m), 3.83 (1H, d, J = 18.9 Hz), 3.77(1H, d, J = 18.9 Hz), 3.62-3.56 (1H, m), 3.00-2.91 (1H, m), 2.40- 2.29(1H, m), 2.27-2.20 (1H, m), 1.99-1.91 (1H, m), 1.63-1.49 (1H, m). 38

Racemate 1H-NMR (DMSO-D6) δ: 11.56 (1H, br s), 8.06 (1H, s), 7.11-7.08(1H, m), 6.55 (1H. dd, J = 3.5, 1.9 Hz), 4.18- 4.07 (2H, m), 3.97-3.85(2H, m), 3.71 (2H, s), 3.65 (2H, s), 2.43-2.22 (2H, m), 1.24 (3H, s),1.20 (3H, s). 39

Racemate 1H-NMR (CDCl3) δ: 10.34 (1H, br s), 8.29 (1H, s), 7.06 (1H, d,J = 3.5 Hz), 6.52 (1H, d, J = 3.7 Hz), 4.76- 4.68 (1H, m), 4.59 (1H, d,J = 13.0 Hz), 4.11 (1H, d, J = 12.8 Hz), 3.75- 3.70 (2H, m), 3.66-3.60(1H, m), 3.54-3.47 (1H, m), 3.39 (3H, s), 3.22- 3.12 (2H, m), 2.64-2.51(2H, m), 2.12-2.04 (1H, m), 1.96-1.61 (5H, m), 1.59-1.52 (1H, m). 40

Single diastereomer(Optically- active substance) 1H-NMR (DMSO-D6) δ:11.73 (1H, br s), 8.14 (1H, s), 7.17 (1H, dd, J = 3.5, 2.3 Hz), 6.61(1H, dd, J = 3.5, 1.6 Hz), 4.47-4.37 (1H, m), 4.30-4.24 (1H, m),4.22-4.05 (2H, m), 4.12 (1H, d, J = 19.2 Hz), 4.04 (1H, d, J = 19.0 Hz),3.65-3.60 (1H, m), 3.54- 3.48 (1H, m), 2.70-2.64 (1H, m), 2.08-1.97 (1H,m), 1.71-1.63 (1H, m), 1.08 (3H, d, J = 7.0 Hz).. 41

Racemate 1H-NMR (DMSO-D6) δ: 11.57 (1H, br s), 8.07 (1H, s), 7.09 (1H,dd, J = 3.5, 2.4 Hz), 6.56 (1H, dd, J = 3.5, 2.0 Hz), 4.01-3.81 (4H, m),3.96 (1H, d, J = 18.7 Hz), 3.90 (1H, d, J = 19.0 Hz), 3.48-3.42 (2H, m),2.46-2.37 (1H, m), 2.16-2.06 (1H, m), 1.77- 1.69 (2H, m), 1.03 (3H, s),0.98 (3H, s). 42

Diastereomer of Compound 33 (Optically-active substance) 1H-NMR(DMSO-D6) δ: 11.57 (1H, br s), 8.08 (1H, s), 7.11-7.09 (1H, m), 6.50(1H, dd, J = 3.5, 1.6 Hz), 4.73- 4.65 (1H, m), 4.29-4.23 (1H, m), 3.94(2H, s), 3.72-3.66 (1H, m), 3.51- 3.44 (1H, m), 3.43-3.36 (1H, m),3.23-3.15 (1H, m), 2.28-2.21 (1H, m), 2.13-2.05 (1H, m), 1.94-1.86 (2H,m), 1.60-1.54 (1H, m), 1.31 (3H, d, J = 6.5 Hz). 43

Diastereomer mixture. Configuration of α- position of carbonyl of amideis S. 1H-NMR (CDCl3) δ: 10.97-10.54 (1H, m), 8.30 (1H, s), 7.11-7.08(1H, m), 6.51 (1H, d, J = 3.2 Hz), 4.80- 4.59 (2H, m), 4.36-4.24 (1H,m), 4.12-4.01 (1H, m), 3.89 (1H, br s), 3.71-3.65 (0.5H, m), 3.58-3.51(1H, m), 3.45-3.37 (0.5H, m), 3.29-3.05 (2H, m), 2.21-2.14 (0.5H, m),2.11- 2.04 (0.5H, m), 2.02-1.70 (5H, m), 1.66-1.53 (1H, m), 1.39-1.32(3H, m). 44

Racemate 1H-NMR (CDC13) δ: 10.67-10.25 (1H, m), 8.30 (1H, s), 7.11-7.07(1H, m), 6.52 (1H, d, J = 3.2 Hz), 4.78- 4.71 (1H, m), 4.70-4.64 (1H,m), 4.09 (1H, d, J = 15.3 Hz), 4.04-3.98 (1H, m), 4.03 (1H, d, J = 15.3Hz), 3.67 (1H, br s), 3.50-3.42 (1H, m), 3.41-3.32 (1H, m), 3.26-3.17(1H, m), 3.17-3.08 (1H, m), 2.18-2.10 (1H, m), 2.04-1.58 (6H, m). 45

Diastereomer mixture. Configuration of α- position of carbonyl of amideis S (derived from (S)-(−)-2- methoxypropionic acid). 1H-NMR (CDCl3) δ:10.62 (1H, br s), 8.30 (1H, s), 7.08 (1H, d, J = 3.5 Hz), 6.54-6.50 (1H,m), 4.75-4.57 (2H, m), 4.20-4.12 (1H, m), 4.05- 3.99 (1H, m), 3.85-3.78(0.5H, m), 3.67-3.62 (1H, m), 3.54-3.46 (0.5H, m), 3.37-3.35 (3H, m),3.27-3.12 (2H, m), 2.13-2.04 (1H, m), 1.99- 1.49 (6H, m), 1.41-1.36 (3H,m). 46

Diastereomer mixture. Configuration of α- position of carbonyl of amideis R (derived from (R)-(+)-2- methoxypropionic acid). 1H-NMR (CDCl3) δ:10.98 (1H, br s), 8.30 (1H, s), 7.09 (1H, d, J = 3.5 Hz), 6.53-6.50 (1H,m), 4.74-4.58 (2H, m), 4.20-4.12 (1H, m), 4.05- 3.99 (1H, m), 3.85-3.78(0.5H, m), 3.68-3.62 (1H, m), 3.53-3.46 (0.5H, m), 3.37-3.35 (3H, m),3.27-3.14 (2H, m), 2.13-2.04 (1H, m), 1.99- 1.48 (6H, m), 1.40-1.36 (3H,m). 47

Diastereomer mixture. Configuration of α- position of carbonyl of amideis S (derived from (S)-(−)-2- acetoxypropionyl chloride). 1H-NMR(DMSO-D6) δ: 11.67 (1H, br s), 8.11 (0.5H, s), 8.10 (0.5H, s), 7.18-7.16(1H, m), 6.58 (0.5H, dd, J = 3.5, 1.8 Hz), 6.51 (0.5H, dd, J = 3.7, 1.8Hz), 5.10-5.03 (1H, m), 4.70- 4.60 (1H, m), 4.52-4.45 (1H, m), 3.88-3.79(1H, m), 3.77-3.70 (0.5H, m), 3.61-3.51 (1H, m), 3.49-3.41 (0.5H, m),3.01-2.82 (2H, m), 2.05 (1.5H, s), 2.04 (1.5H, s), 1.91-1.72 (4H, m),1.66-1.56 (2H, m), 1.50- 1.43 (1H, m), 1.35 (1.5H, d, J = 6.8 Hz), 1.31(1.5H, d, J = 6.8 Hz). 48

Racemate 1H-NMR (DMSO-D6) δ: 11.67 (1H, br s), 8.08 (1H, s), 7.15 (1H,dd, J = 3.5, 2.3 Hz), 6.56 (1H, dd, J = 3.6, 1.7 Hz), 4.72-4.66 (1H, m),4.48 (1H, d, J = 12.5 Hz), 3.98 (1H, d, J = 14.6 Hz), 3.94 (1H, d, J =14.8 Hz), 3.89 (1H, d, J = 13.0 Hz), 3.53-3.46 (1H, m), 3.39-3.34 (1H,m), 3.30 (3H, s), 2.99-2.89 (2H, m), 1.85-1.66 (4H, m), 1.62-1.52 (2H,m), 1.51-1.45 (1H, m). 49

Racemate 1H-NMR (DMSO-D6) δ: 11.67 (1H, br s), 8.08 (1H, s), 7.13 (1H,dd, J = 3.5, 2.6 Hz), 6.54 (1H, dd, J = 3.6, 1.7 Hz), 5.97 (1H, t, J =5.6 Hz), 4.72-4.64 (1H, m), 4.40 (1H, d, J = 12.5 Hz), 3.87 (1H, d, J =12.5 Hz), 3.39-3.29 (1H, m), 3.28-3.21 (1H, m), 3.07-3.00 (2H, m),2.96-2.84 (2H, m), 1.83-1.65 (4H, m), 1.62- 1.47 (2H, m), 1.44-1.37 (1H,m), 1.02 (3H, t, J = 7.1 Hz). 50

Racemate 1H-NMR (CDCl3) δ: 10.68 (1H, br s), 8.28 (1H, s), 7.07 (1H, d,J = 3.2 Hz), 6.51 (1H, d, J = 3.7 Hz), 4.75- 4.68 (1H, m), 4.67-4.61(1H, m), 4.64 (1H, d, J = 14.6 Hz), 4.60 (1H, d, J = 14.8 Hz), 4.06-4.01(1H, m), 3.58-3.51 (1H, m), 3.49-3.42 (1H, m), 3.21-3.07 (2H, m), 2.21(3H, s), 2.13-2.05 (1H, m), 2.03-1.81 (3H, m), 1.79-1.55 (3H, m). 51

Racemate 1H-NMR (CDCl3) δ: 9.34 (1H, br s), 8.28 (1H, s), 7.00-6.97 (1H,m), 6.58-6.54 (1H, m), 4.19-3.95 (2H, m), 3.84-3.45 (6H, m), 3.24-3.08(2H, m), 2.28-2.14 (2H, m), 2.07- 1.90 (5H, m), 1.85-1.61 (2H, m). 52

Racemate 1H-NMR (DMSO-D6) δ: 11.68 (1H, br s), 8.10 (1H, s), 7.16 (1H,dd, J = 3.5, 2.6 Hz), 6.82 (1H, t, J = 5.6 Hz), 6.59 (1H, dd, J = 3.7,1.9 Hz), 4.75- 4.67 (1H, m), 4.51-4.45 (1H, m), 4.03 (2H, d, J = 5.8Hz), 3.86-3.81 (1H, m), 3.44-3.37 (1H, m), 3.32- 3.25 (1H, m), 2.97-2.85(2H, m), 1.88-1.69 (4H, m), 1.66-1.52 (2H, m), 1.52-1.45 (1H, m). 53

Racemate 1H-NMR (CDCl3) δ: 10.30-10.10 (1H, m), 8.33 (1H, s), 7.06 (1H,d, J = 2.8 Hz), 6.60 (1H, d, J = 3.0 Hz), 4.10-4.03 (1H, 8.10 (1H, m),3.87- 3.62 (3H, m), 2.53 (2H, t, J = 6.7 Hz), 2.22-2.08 (1H, m),1.99-1.85 (6H, m) 54

Single diastereomer (Optically-active substance) 1H-NMR (DMSO-D6) δ:12.75 (1H, br s), 8.32 (1H, s), 7.49-7.45 (1H, m), 6.98-6.93 (1H, m),4.26-3.78 (5H, m), 3.97 (2H, s), 3.58-3.23 (2H, m), 2.83-2.73 (1H, m),2.28-2.20 (1H, m), 2.17-1.96 (2H, m), 1.70-1.49 (1H, m), 1.02 (3H, d, J= 6.7 Hz). 55

Racemate 1H-NMR (DMSO-D6) δ: 11.70 (1H, br s), 8.14 (1H, s), 7.18 (1H,dd, J = 3.5, 2.3 Hz), 6.63 (1H, dd, J = 3.5, 1.9 Hz), 4.22-4.01 (5H, m),3.96-3.82 (1H, m), 3.72-3.60 (2H, m), 2.86- 2.77 (1H, m), 2.72-2.51 (2H,m), 2.41-2.30 (1H, m). 56

Racemate 1H-NMR (DMSO-D6) δ: 11.67 (1H, br s), 8.11 (1H, s), 7.17 (1H,dd, J = 3.5, 2.6 Hz), 6.61 (1H, dd, J = 3.7, 1.9 Hz), 4.90-4.84 (1H, m),4.63-4.56 (1H, m), 4.06 (1H, d, J = 19.0 Hz), 3.99 (1H, d, J = 19.0 Hz),3.94-3.88 (1H, m), 3.67-3.60 (1H, m), 3.56- 3.49 (1H, m), 3.16-3.08 (1H,m), 2.81-2.71 (1H, m), 2.48-2.37 (2H, m), 2.00-1.90 (2H, m), 1.85-1.72(1H, m). 57

Racemate 1H-NMR (DMSO-D6) δ: 11.59 (1H, br s), 8.08 (1H, s), 7.13-7.10(1H, m), 6.55 (1H, dd, J = 3.2, 1.6 Hz), 4.33- 4.28 (1H, m), 4.08 (2H,s), 3.97-3.54 (3H, m), 3.48-3.29 (2H, m), 2.61- 2.53 (1H, m), 2.30-2.06(1H, m), 1.82-1.47 (6H, m). 58

Racemate 1H-NMR (DMSO-D6) δ: 11.67 (1H, br s), 8.10 (1H, s), 7.16 (1H,dd, J = 3.5, 2.6 Hz), 6.56 (1H, dd, J = 3.6, 1.7 Hz), 6.46 (1H, t, J =5.6 Hz), 4.74-4.67 (1H, m), 4.45 (1H, d, J = 12.5 Hz), 3.87 (1H, d, J =12.8 Hz), 3.43-3.36 (1H, m), 3.30-3.24 (3H, m), 2.96-2.87 (2H, m), 2.66(2H, t, J = 6.6 Hz), 1.87-1.68 (4H, m), 1.65- 1.51 (2H, m), 1.49-1.42(1H, m). 59

Racemate of Compound 3 1H-NMR (DMSO-D6) δ: 11.71 (1H, br s), 8.13 (1H,s), 7.20 (1H, dd, J = 3.5, 2.4 Hz), 6.64 (1H, dd, J = 3.6, 1.9 Hz), 4.96(1H, d, J = 12.6 Hz), 4.67-4.60 (1H, m), 4.11-4.04 (1H, m), 4.00-3.93(1H, m), 3.70 (2H, s), 3.53 (1H, d, J = 12.6 Hz), 2.98-2.90 (1H, m),2.39-2.31 (1H, m), 2.01- 1.93 (3H, m), 1.83-1.75 (1H, m), 1.58-1.46 (1H,m). 60

Single diastereomer (Optically-active substance) 1H-NMR (DMSO-D6) δ:11.67 (1H, br s), 8.12 (1H, s), 7.16 (1H, dd, J = 3.5, 2.6 Hz), 6.61(1H, dd, J = 3.7, 1.9 Hz), 4.60 (1H, d, J = 13.2 Hz), 4.38-4.30 (1H, m),3.98 (1H, d, J = 18.8 Hz), 3.92 (1H, d, J = 19.0 Hz), 3.75 (1H, d, J =13.2 Hz), 3.54-3.47 (1H, m), 3.46-3.35 (1H, m), 3.28- 3.20 (1H, m),2.54-2.46 (1H, m), 2.18-2.07 (2H, m), 2.01-1.92 (1H, m), 1.87-1.76 (1H,m), 1.66-1.58 (1H, m), 1.56-1.47 (1H, m), 0.82 (3H, d, J = 7.0 Hz). 61

Racemate 1H-NMR (CDCl3) δ: 10.99-10.59 (1H, m), 8.31 (1H, s), 7.08-7.01(1H, m), 6.55 (1H, d, J = 3.0 Hz), 4.63- 4.03 (2H, m), 3.84-3.44 (4H,m), 3.70 (3H, s), 3.31-3.12 (0.7H, m), 3.00-2.81 (0.3H, m), 2.16-1.71(5H, m). 62

Racemate 1H-NMR (DMSO-D6) δ: 11.56 (1H, br s), 8.07 (1H, s), 7.09 (1H,dd, J = 3.2, 2.6 Hz), 6.92 (1H, t, J = 5.7 Hz), 6.55 (1H, dd, J = 3.2,1.9 Hz), 4.37- 4.12 (1H, m), 4.05-3.93 (1H, m), 4.02 (2H, d, J = 5.6Hz), 3.79-3.49 (2H, m), 3.40-3.27 (2H, m), 3.07- 2.94 (1H, m), 1.98-1.83(4H, m), 1.76-1.67 (1H, m). 63

Racemate 1H-NMR (CDCl3) δ: 9.13 (1H, br s), 8.29 (1H, s), 7.00-6.97 (1H,m), 6.58-6.54 (1H, m), 4.18-4.09 (1H, m), 4.06-3.93 (2H, m), 3.88-3.72(1H, m), 3.68-3.61 (1H, m), 3.59- 3.53 (2H, m), 3.22-3.09 (1H, m), 2.98(3H, s), 2.07-1.92 (5H, m), 1.87- 1.69 (1H, m). 64

Racemate 1H-NMR (CDCl3) δ: 9.33-9.10 (1H, m), 8.32 (1H, s), 7.79 (1H,s), 7.76- 7.70 (2H, m), 7.58-7.52 (1H, m), 7.03-7.00 (1H, m), 6.61-6.57(1H, m), 4.75-4.55 (1H, m), 4.27-4.18 (1H, m), 3.97-3.81 (1H, m), 3.79-3.72 (1H, m), 3.57-3.40 (3H, m), 2.23-2.15 (1H, m), 2.12-2.03 (1H, m),1.98-1.85 (3H, m). 65

Racemate 1H-NMR (CDCl3) δ: 9.44 (1H, br s), 8.32 (1H, s), 7.73 (2H, d, J= 8.3 Hz), 7.58 (2H, d, J = 8.1 Hz), 7.02 (1H, dd, J = 3.6, 2.2 Hz),6.59 (1H, dd, J = 3.2, 1.9 Hz), 4.75-4.52 (1H, m), 4.29-4.17 (1H, m),3.97-3.82 (1H, m), 3.78-3.73 (1H, m), 3.53- 3.41 (3H, m), 2.22-2.15 (1H,m), 2.11-2.03 (1H, m), 1.97-1.87 (3H, m). 66

Single diastereomer (Optically-active substance) 1H-NMR (DMSO-D6) δ:11.60 (1H, br s), 8.08 (1H, s), 7.11 (1H, dd, J = 3.5, 2.6 Hz), 6.58(1H, dd, J = 3.5, 1.9 Hz), 3.99 (1H, d, J = 19.0 Hz), 3.98-3.78 (4H, m),3.92 (1H, d, J = 19.2 Hz), 3.54-3.47 (1H, m), 3.44-3.38 (1H, m),3.05-2.95 (1H, m), 2.21-2.11 (1H, m), 1.98-1.87 (2H, m), 1.63-1.52 (1H,m), 0.93 (3H, d, J = 6.7 Hz). 67

Racemate 1H-NMR (DMSO-D6) δ: 11.55 (1H, br s), 8.05 (1H, s), 7.08 (1H,dd, J = 3.2, 2.6 Hz), 6.58-6.51 (2H, m), 4.35-4.16 (1H, m), 4.00-3.92(1H, m), 3.72-3.50 (2H, m), 3.39-3.27 (2H, m), 3.26-3.21 (2H, m), 3.08-2.95 (1H, m), 2.62 (2H, t, J = 6.6 Hz), 1.94-1.82 (4H, m), 1.71-1.63(1H, m). 68

Racemate 1H-NMR (DMSO-D6) δ: 11.55 (1H, s), 8.06 (1H, s), 7.08 (1H, dd,J = 3.4, 2.4 Hz), 6.55-6.52 (1H, m), 6.10 (1H, t, J = 5.6 Hz), 4.37-4.15(1H, m), 4.01-3.90 (1H, m), 3.69- 3.55 (1H, m), 3.54-3.49 (1H, m),3.37-3.28 (2H, m), 3.09-3.01 (1H, m), 2.99-2.93 (2H, m), 1.93-1.80 (4H,m), 1.68-1.62 (1H, m), 1.46- 1.37 (2H, m), 0.83 (3H, t, J = 7.4 Hz). 69

Racemate 1H-NMR (DMSO-D6) δ: 11.58 (1H, br s), 8.06 (1H, s), 7.10-7.07(1H, m), 6.55-6.49 (1H, m), 4.39-4.10 (1H, m), 4.04-3.92 (1H, m),3.83-3.62 (1H, m), 3.62-3.49 (3H, m), 3.56 (3H, s), 3.07-2.91 (1H, m),2.60-2.53 (2H, m), 2.51-2.45 (2H, m), 1.99- 1.93 (2H, m), 1.91-1.82 (2H,m), 1.75-1.65 (1H, m). 70

Racemate 1H-NMR (CDCl3) δ: 10.81 (1H, br s), 8.29 (1H, s), 7.02 (1H, d,J = 3.7 Hz), 6.52 (1H, d, J = 3.5 Hz), 4.64 (2H, s), 4.55-4.39 (1H, m),4.18-4.07 (1H, m), 3.85-3.74 (1H, m), 3.72- 3.64 (1H, m), 3.59-3.48 (2H,m), 3.37-3.26 (1H, m), 2.19 (3H, s), 2.14- 1.91 (4H, m), 1.83-1.72 (1H,m). 71

Racemate 1H-NMR (DMSO-D6) δ: 11.58 (1H, br s), 8.27 (2H, d, J = 9.0 Hz),8.07 (1H, s), 7.46 (2H, d, J = 9.3 Hz), 7.12-7.09 (1H, m), 6.60-6.57(1H, m), 4.29-4.15 (1H, m), 4.05-3.95 (1H, m), 3.78-3.65 (4H, m), 2.98-2.82 (1H, m), 2.15-2.03 (2H, m), 1.97-1.86 (3H, m). 72

Racemate 1H-NMR (DMSO-D6) δ: 11.57 (1H, br s), 8.07 (1H, s), 7.10-7.08(1H, m), 6.56-6.52 (1H, m), 4.46 (1H, t, J = 5.6 Hz), 4.39-4.21 (1H, m)4.06-3.96 (3H, m), 3.74-3.55 (2H, m), 3.45- 3.39 (2H, m), 3.09-2.97 (1H,m), 1.97-1.92 (2H, m), 1.90-1.84 (2H, m), 1.78-1.71 (1H, m). 73

Racemate 1H-NMR (DMSO-D6) δ: 11.58 (1H, br s), 8.06 (1H, s), 7.10 (1H,dd, J = 3.4, 2.4 Hz), 6.57 (1H, dd, J = 3.5, 2.1 Hz), 4.30-4.09 (1H, m),4.04-3.94 (1H, m), 3.92-3.82 (2H, m), 3.77- 3.52 (2H, m), 2.97-2.84 (1H,m), 2.07-1.92 (4H, m), 1.85-1.76 (1H, m), 1.63-1.49 (4H, m). 74

Racemate 1H-NMR (DMSO-D6) δ: 11.63 (1H, s), 8.09 (1H, s), 7.15 (1H, dd.J = 3.5, 2.6 Hz), 6.64 (1H, dd, J = 3.6, 2.0 Hz), 4.53 (1H, d, J = 13.2Hz), 4.38- 4.29 (1H, m), 4.18 (1H, d, J = 13.0 Hz), 4.02 (2H, s),3.35-3.31 (3H, m), 2.63-2.55 (1H, m), 1.88-1.77 (2H, m), 1.74-1.50 (6H,m), 1.47-1.34 (3H, m). 75

Racemate 1H-NMR (DMSO-D6) δ: 11.59 (1H, br s), 8.08 (1H, s), 7.13 (1H,dd, J = 3.5, 2.6 Hz), 6.57 (1H, dd, J = 3.6, 1.7 Hz), 3.90-3.83 (1H, m),3.70-3.62 (1H, m), 3.69 (1H, d, J = 12.8 Hz), 3.53 (1H, d, J = 12.8 Hz),2.67-2.57 (1H, m), 2.54-2.45 (1H, m), 1.76- 1.63 (2H, m), 1.60-1.36(10H, m). 76

Racemate 1H-NMR (DMSO-D6) δ: 11.67 (1H, br s), 8.08 (1H, s), 7.18-7.14(1H, m), 6.69-6.63 (1H, m), 4.53-4.45 (1H, m), 4.33-4.24 (1H, m),4.14-4.07 (1H, m), 3.45-3.25 (3H, m), 2.52- 2.39 (1H, m), 1.85-1.47(11H, m), 1.40 (9H, s). 77

Racemate 1H-NMR (DMSO-D6) δ: 11.60 (1H, br s), 8.08 (1H, s), 7.13-7.10(1H, m), 6.57-6.51 (1H, m), 4.32-4.26 (1H, m), 4.07 (2H, s), 4.00-3.50(3H, m), 3.48-3.32 (2H, m), 2.60-2.52 (1H, m), 2.30-2.08 (1H, m),1.82-1.44 (6H, m). 78

Racemate 1H-NMR (DMSO-D6) δ: 11.59 (1H, br s), 8.08 (1H, s), 7.13-7.11(1H, m), 6.55-6.52 (1H, m), 4.39-4.33 (1H, m), 3.92-3.74 (2H, m),3.64-3.51 (2H, m), 3.42-3.36 (1H, m), 2.65- 2.56 (1H, m), 2.24-1.97 (1H,m), 2.06 (3H, s), 1.87-1.73 (1H, m), 1.71- 1.56 (4H, m), 1.52-1.40 (1H,m). 79

Racemate 1H-NMR (DMSO-D6) δ: 11.53 (1H, br s), 8.05 (1H, s), 7.08 (1H,d, J = 3.5 Hz), 6.55 (1H, t, J = 4.4 Hz), 3.91-3.67 (3H, m), 3.61-3.40(1H, m), 2.76-2.62 (2H, m), 2.04-1.91 (2H, m), 1.88-1.79 (1H, m), 1.64-1.46 (4H, m), 1.45-1.37 (2H, m). 80

Racemate 1H-NMR (DMSO-D6) δ: 11.58 (1H, br s), 8.07 (1H, s), 7.12-7.10(1H, m), 6.55 (1H, dd, J = 3.2, 1.9 Hz), 4.31- 4.21 (1H, m), 4.00-3.47(4H, m), 3.33-3.22 (1H, m), 2.59-2.44 (1H, m), 2.24-2.04 (1H, m),1.82-1.71 (1H, m), 1.69-1.56 (3H, m), 1.53- 1.45 (1H, m), 1.45-1.36 (1H,m), 1.42 (9H, s). 81

Racemate 1H-NMR (DMSO-D6) δ: 12.02 (1H, br s), 8.17 (1H, s), 7.27-7.23(1H, m), 6.75-6.69 (1H, in), 4.57-4.35 (3H, m), 3.46-3.33 (3H, m),2.78-2.68 (1H, m), 2.02 (3H, s), 1.87-1.78 (1H, m), 1.75-1.40 (8H, m).82

Racemate 1H-NMR (DMSO-D6) δ: 11.68 (1H, br s), 8.10 (1H, s), 7.17 (1H,dd, J = 3.5, 2.6 Hz), 6.59 (1H, dd, J = 3.7, 1.9 Hz), 4.64-4.59 (1H, m),4.51-4.44 (1H, m), 4.26-4.21 (1H, m), 4.07 (1H, d, J = 18.8 Hz), 4.01(1H, d, J = 18.6 Hz), 3.31-3.13 (2H, m), 2.77- 2.67 (1H, m), 1.86-1.37(10H, m). 83

Racemate 1H-NMR (DMSO-D6) δ: 11.65 (1H, br s), 8.09 (1H, s), 7.15 (1H,dd, J = 3.4, 2.5 Hz), 6.57 (1H, dd, J = 3.6, 1.9 Hz), 4.73-4.65 (1H, m),4.47 (1H, d, J = 12.1 Hz), 3.90 (1H, d, J = 12.4 Hz), 3.62-3.54 (1H, m),3.48-3.41 (1H, m), 3.00-2.88 (2H, m), 1.97 (3H, s), 1.88-1.67 (4H, m),1.65-1.53 (2H, m), 1.50-1.43 (1H, m). 84

Racemate of Compound 7 1H-NMR (DMSO-D6) δ: 11.64 (1H, br s), 8.10 (1H,s), 7.16 (1H, dd, J = 3.4, 2.6 Hz), 6.57 (1H, dd, J = 3.6, 1.6 Hz),4.73-4.66 (1H, m), 4.52 (1H, d, J = 12.5 Hz), 3.93 (2H, s), 3.83 (1H, d,J = 12.9 Hz), 3.58-3.51 (1H, m), 3.46-3.39 (1H, m), 2.99-2.84 (2H, m),1.90-1.70 (4H, m), 1.67- 1.57 (2H, m), 1.56-1.49 (1H, m). 85

Racemate 1H-NMR (CDCl3) δ: 9.05 (1H, br s), 8.27 (1H, s), 6.96 (1H, d, J= 3.7 Hz), 6.55 (1H, d, J = 3.5 Hz), 4.19- 4.10 (1H, m), 3.90-3.75 (1H,m), 3.70-3.56 (3H, m), 3.39-3.28 (1H, m), 2.70-2.60 (1H, m), 2.12-2.03(1H, m), 1.99-1.91 (4H, m), 1.79- 1.69 (1H, m), 1.16-1.08 (6H, m). 86

Racemate 1H-NMR (DMSO-D6) δ: 11.56 (1H, br s), 8.07 (1H, s), 7.10-7.07(1H, m), 6.56-6.51 (1H, m), 4.36-4.22 (1H, m), 4.04-3.94 (1H, m),3.72-3.59 (1H, m), 3.56-3.49 (3H, m), 3.11- 2.97 (1H, m), 2.29 (2H, q, J= 7.5 Hz), 1.97-1.93 (2H, m), 1.90-1.82 (2H, m), 1.74-1.66 (1H, m), 0.97(3H, t, J = 7.4 Hz). 87

Racemate 1H-NMR (DMSO-D6) δ: 11.58 (1H, br s), 8.07 (1H, s), 7.11-7.08(1H, m), 6.57-6.52 (1H, m), 4.38-4.24 (1H, m), 4.03-3.94 (1H, m),3.75-3.60 (1H, m), 3.59-3.48 (3H, m), 3.13- 2.99 (1H, m), 1.99 (3H, s),1.98-1.93 (2H, m), 1.90-1.82 (2H, m), 1.74- 1.66 (1H, m). 88

Racemate 1H-NMR (DMSO-D6) δ: 11.59 (1H, br s), 8.08 (1H, s), 7.10 (1H,dd, J = 3.4, 2.4 Hz), 6.56-6.52 (1H, m), 4.36- 4.17 (1H, m), 4.10-3.94(1H, m), 3.99 (2H, s), 3.76-3.19 (4H, m), 3.05- 2.92 (1H, m), 2.01-1.94(2H, m), 1.93-1.83 (2H, m), 1.81-1.72 (1H, m). 89

Racemate 1H-NMR (CDCl3) δ: 11.52 (1H, s), 8.05 (1H, s), 7.08 (1H, dd, J= 3.2, 1.9 Hz), 6.55 (1H, d, J = 2.3 Hz), 3.87-3.71 (2H, m), 3.67-3.53(2H, m), 2.91-2.81 (2H, m), 2.31 (1H, br s), 1.99-1.84 (2H, m),1.81-1.63 (4H, m). 90

Racemate 1H-NMR (DMSO-D6) δ: 11.56 (1H, br s), 8.07 (1H, s), 7.35-7.27(4H, m), 7.22-7.18 (1H, m), 7.11-7.09 (1H, m), 6.62-6.58 (1H, m),4.04-3.91 (1H, m), 3.80-3.56 (5H, m), 2.68- 2.55 (2H, m), 2.29-2.19 (1H,m), 1.91-1.71 (5H, m).

TABLE 2 No. Structural Formula Comments NMR Data 91

Racemate 1H-NMR (DMSO-D6) δ: 13.60 (1H, br s), 8.24 (1H, s), 8.17 (1H,s), 4.02- 3.98 (1H, m), 3.93 (2H, s), 3.81-3.75 (1H, m), 3.51-3.38 (2H,m), 3.27- 3.05 (2H, m), 2.94-2.84 (1H, m), 2.37-2.28 (1H, m), 1.85-1.69(5H, m), 1.57-1.50 (1H, m). 92

Racemate 1H-NMR (DMSO-D6) δ: 13.32 (1H, br s), 8.18 (1H, s), 8.01 (1H,s), 4.66- 4.62 (1H, m), 4.09-4.00 (2H, m), 3.99 (2H, s), 3.60-3.56 (1H,m), 3.53- 3.42 (2H, m), 3.06-2.97 (1H, m), 2.03-1.81 (4H, m), 1.77-1.71(1H, m). 93

Racemate 1H-NMR (DMSO-D6) δ: 13.32 (1H, br s), 8.13 (1H, d, J = 1.4 Hz),8.02 (1H, s), 4.39 (1H, d, J = 10.9 Hz), 4.28 (1H, d, J = 10.7 Hz), 4.09(2H, s), 4.08-4.01 (1H, m), 3.88-3.82 (1H, m), 3.50-3.43 (1H, m),3.38-3.27 (1H, m), 2.61-2.54 (1H, m), 2.16- 2.06 (1H, m), 1.84-1.45 (6H,m).

TABLE 3 No. Structural Formula Comments NMR Data 94

Racemate 1H-NMR (DMSO-D6) δ: 13.02 (1H, br s), 8.17 (1H, s), 8.11 (1H,s), 5.72-4.95 (1H, m), 4.05 (1H, d, J = 19.2 Hz), 3.94 (1H, d, J = 19.0Hz), 3.89-3.78 (1H, m), 3.60-3.52 (1H, m), 3.46-3.39 (1H, m), 2.97-2.73(2H, m), 1.94-1.84 (2H, m), 1.82-1.49 (6H, m). 95

Racemate 1H-NMR (DMSO-D6) δ: 12.92 (1H, br s), 8.17 (1H, s), 8.07 (1H,s), 4.95-4.78 (1H, m), 4.34-4.09 (1H, m), 4.06 (2H, s), 3.99-3.67 (1H,m), 3.60-3.50 (1H, m), 3.43-3.28 (2H, m), 2.58-2.41 (1H, m), 2.30-2.11(1H, m), 1.81-1.48 (6H, m).

The following Table 4 shows chemical structures of compounds of whichabsolute configurations have been specified among the aboveoptically-active compounds.

TABLE 4 No. Structural Formula  3

 4

 9

25

[Test 1]

JAK3 activity inhibitory effects of test compounds were evaluated by thefollowing kinase reactions.

In the kinase reactions, fused proteins (6His tag-fused hJAK3 kinasedomain (aa781-end)) which were coexpressed in Sf21 cells and purified byNi2+/NTA agarose were used. The kinase reactions were initiated by theaddition of the following solutions of (a) to (c) to 96-well half-areawhite plates (plates, Corning Incorporated 3642).

(a) 5 μmol/L TK substrate-biotin (cisbio) diluted by kinase buffer (50mmol/L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.0)),0.02% sodium azide, 0.1 mmol/L sodium vanadate, 5 mmol/L magnesiumchloride, 1 mmol/L dithiothreitol, 0.01% bovine serum albumin), 25μmol/L ATP, 250 nmol/L Supplement Enzymatic buffer (cisbio) solution: 10μL/well(b) Test-article solution prepared by using kinase buffer containing 5%dimethylsulfoxide: 10 μL/well(c) 33 ng/mL hJAK3 enzyme diluted by kinase buffer: 30 μL/well

A well in which ATP was not added was set out as a blank well.

Plates were let stand at room temperature for 10 minutes from startingreactions.

To the plates were added 50 μL/well of a buffer for detection containingTK-Antibody-Cryptate (5 tests/50 μL) and streptoavidine-addition XL665(62.5 nmol/L) reagent (50 mmol/L4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.0), 20 mM EDTA,800 mmol/L potassium fluoride, 0.1% bovine serum albumin).

One hour after the addition of the buffer for detection, fluorescencecounts of each well were measured by a fluorescence microplate reader.Specifically, fluorescence counts in 620 nm excited in 337 nm, andfluorescence counts in 665 nm excited by fluorescence in 620 nm weremeasured.

Ratio of each well was calculated from measured fluorescence counts(fluorescence counts in 665 nm/fluorescence counts in 620 nm×10000).

Data were obtained by deducting the average Ratio of a blank well fromRatio of each well. IC50 values of test articles were calculated from %of control values of 2 doses before as well as after 50% in 100% as % ofa control value of a solvent control. % Inhibition of either 0.1 μmol/Lor 1 μmol/L (100-% of control values) was also calculated.

[Test 2]

JAK2 activity inhibitory effects of test compounds were evaluated by thefollowing kinase reactions.

In the kinase reactions, fused proteins (6His tag-fused hJAK2 kinasedomain (aa808-end)) which were coexpressed in Sf21 cells and purified byNi2+/NTA agarose were used. The kinase reactions were initiated by theaddition of the following solutions of (a) to (c) to 96-well half-areawhite plates (plates, Corning Incorporated 3642).

(a) 5 μmol/L TK substrate-biotin (cisbio) diluted by kinase buffer (50mmol/L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.0)),0.02% sodium azide, 0.1 mmol/L sodium vanadate, 5 mmol/L magnesiumchloride, 1 mmol/L dithiothreitol, 0.01% bovine serum albumin), 100mol/L ATP, 250 nmol/L Supplement Enzymatic buffer (cisbio) solution: 10μL/well(b) Test-article solution prepared by using kinase buffer containing 5%dimethylsulfoxide: 10 μL/well(c) 7 ng/mL hJAK2 enzyme diluted by kinase buffer: 30 μL/well

A well in which ATP was not added was set out as a blank well.

Plates were let stand at room temperature for 10 minutes from startingreactions.

To the plates were added 50 μL/well of a buffer for detection containingTK-Antibody-Cryptate (5 tests/50 μL) and streptoavidine-addition XL665(62.5 nmol/L) reagent (50 mmol/L4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.0), 20 mM EDTA,800 mmol/L potassium fluoride, 0.1% bovine serum albumin).

One hour after the addition of the buffer for detection, fluorescencecounts of each well were measured by a fluorescence microplate reader.Specifically, fluorescence counts in 620 nm excited in 337 nm, andfluorescence counts in 665 nm excited by fluorescence in 620 nm weremeasured.

Ratio of each well was calculated from measured fluorescence counts(fluorescence counts in 665 nm/fluorescence counts in 620 nm×10000).

Data were obtained by deducting the average Ratio of a blank well fromRatio of each well. IC50 values of test articles were calculated from %of control values of 2 doses before as well as after 50% in 100% as % ofa control value of a solvent control.

The following Tables 5 to 7 show JAK3 activity inhibition data or %inhibition data of Compounds 1 to 95.

TABLE 5-1 Compound JAK3 % No. IC50 (uM) Inhibition  1 0.0034  2 0.0047 3 0.010  4 0.0051  5 0.0040  6 0.0071  7 0.0058  8 0.73  9 24 (0.1 uM)10 28 (0.1 uM) 11 38 (0.1 uM) 12 46 (0.1 uM) 13 26 (1 uM) 14 0.041 150.0071 16 0.058 17 0.087 18 0.41 19 0.40 20 0.10 21 0.021 22 30 (1 uM)23 0.014 24 0.13 25 0.23 26 0.012 27 0.088 28 0.13 29 0.047 30 0.58 310.046 32 0.49 33 0.48 34 0.036 35 0.53 36 0.15 37 0.0033 38 0.15 39 0.1240 0.14 41 0.090 42 0.10 43 0.040 44 0.067 45 0.27 46 0.27 47 0.11 480.17 49 0.092 50 0.23 51 41 (1 uM) 52 0.010 53 0.056 54 0.0059 55 0.006656 0.0068 57 0.037 58 0.031 59 0.038 60 0.014 61 0.65 62 0.13 63 0.24 640.18 65 0.19 66 0.011 67 0.24 68 0.28 69 0.32 70 0.30 71 0.25 72 0.15 730.073 74 0.18 75 1 (1 uM) 76 0.76 77 0.048 78 0.15 79 38 (1 uM) 80 0.1081 0.46 82 0.013 83 0.25 84 0.014 85 0.57 86 0.45 87 39 (1 uM) 88 0.03289 8.6 90 0.55

TABLE 6 Compound JAK3 No. IC50 (uM) % Inhibition 91 42 (1 uM) 92 0.33 930.11

TABLE 71 Compound JAK3 % No. IC50 (uM) Inhibition 94 0.35 95 0.078

The following Table 8 shows JAK2 activity inhibition data or %inhibition data of Compounds 1 to 7.

TABLE 8 Compound JAK2 % No. IC50 (uM) Inhibition 1 0.0010 2 0.0017 30.0022 4 0.0017 5 0.0019 6 0.0021 7 0.0047

[Formulations]

The formulation examples of the present invention include the followingformulations. However, the present invention is not intended to belimited thereto.

Formulation 1 (Preparation of Capsule)

1) Compound 1 30 mg 2) Microcrystalline cellulose 10 mg 3) Lactose 19 mg4) Magnesium stearate  1 mg

1), 2), 3) and 4) are mixed to fill in a gelatin capsule.

Formulation 2 (Preparation of Tablet)

1) Compound 1 10 g 2) Lactose 50 g 3) Cornstarch 15 g 4) Carmellosecalcium 44 g 5) Magnesium stearate  1 g

The whole amount of 1), 2) and 3) and 30 g of 4) are combined withwater, dried in vacuo, and then granulated. The resulting granules aremixed with 14 g of 4) and 1 g of 5), and tableted by a tabletingmachine. Then, 1000 tablets wherein Compound 1 (10 mg) is comprised ineach tablet are obtained.

INDUSTRIAL APPLICABILITY

The present invention is useful for the treatment or prevention of:

(a) organ transplant rejection, graft versus host reaction aftertransplantation;(b) autoimmune diseases including rheumatoid arthritis, psoriasis,psoriatic arthritis, multiple sclerosis, ulcerative colitis, Crohn'sdisease, systemic lupus erythematosus, type I diabetes, myastheniagravis, Castleman's disease, juvenile idiopathic arthritis, dry eye; and(c) allergic diseases including asthma, atopic dermatitis, rhinitis,etc. The present invention is also useful for the treatment orprevention of chronic myeloproliferative diseases including polycythemiavera, primary myelofibrosis, essential thrombocythemia, etc.

1. A compound of formula [I]:

wherein R^(a) is the same or different and each is: (1) C₁₋₆ alkyl, or(2) halogen atom, n1 is an integer selected from 0 to 4, R^(b) is thesame or different and each is: (1) C₁₋₆ alkyl, or (2) halogen atom, n2is an integer selected from 0 to 4, m1 is an integer selected from 0 to3, m2 is an integer selected from 1 to 4, X^(a)═X^(b) is: (1) CH═CH, (2)N═CH, or (3) CH═N, X is: (1) nitrogen atom, or (2) C—R^(d) wherein R^(d)is hydrogen atom or halogen atom, R^(c) is a group selected from thefollowing (1) to (6): (1) hydrogen atom, (2) C₁₋₆ alkyl optionallysubstituted by the same or different 1 to 5 substituents selected fromthe following Group A, (3) —C(═O)—R^(c1), (4) —C(═O)—O—R^(c2) (5)—C(═O)—NR^(c3)R^(c4) in which R^(c1), R^(c2), R^(c3) and R^(c4) are thesame or different and each is: (i) hydrogen atom, or (ii) C₁₋₆ alkyloptionally substituted by the same or different 1 to 5 substituentsselected from the following Group A, and (6) a group of formula:

in which Y^(a) is a group selected from the following (i) to (iii): (i)C₁₋₆ alkylene, (ii) —C(═O)—, and (iii) —C(═O)—O—, Ring T is: (i) C₆₋₁₀aryl, (ii) C₃₋₁₀ cycloalkyl, or (iii) saturated monoheterocyclyl whereinthe saturated monoheterocyclyl comprises 1 to 4 heteroatoms selectedfrom nitrogen atom, oxygen atom or sulfur atom as well as carbon atomsand the number of the constituent ring atoms is 3 to 7, R^(c5) is thesame or different and each is: (i) cyano, or (ii) nitro, p is an integerselected from 0 to 4, Group A is selected from the group consisting of:(a) hydroxyl, (b) C₁₋₆ alkoxy, (c) cyano, (d) C₁₋₆ alkoxycarbonyl, (e)C₁₋₆ alkylcarbonyloxy, and (f) C₂₋₆ alkenyloxy, or a pharmaceuticallyacceptable salt thereof, or a solvate of the compound or salt thereof,provided that when X^(a)═X^(b) is CH═CH, then X is C—R^(d) wherein R^(d)is hydrogen atom or halogen atom.
 2. The compound as claimed in claim 1,wherein, n1 is an integer selected from 0 to 2, n2 is an integerselected from 0 to 2, m1 is an integer selected from 0 to 3, m2 is aninteger selected from 1 to 3, X is: (1) nitrogen atom, or (2) C—R^(d)wherein R^(d) is halogen atom, R^(c) is a group selected from thefollowing (1) to (6): (1) hydrogen atom, (2) C₁₋₆ alkyl substituted byone substituent selected from the following Group A, (3) —C(═O)—R^(c1),(4) —C(═O)—O—R^(c2), (5) —C(═O)—NR^(c3)R^(c4) in which R^(c1) is C₁₋₆alkyl optionally substituted by one substituent selected from thefollowing Group A, R^(c2) is C₁₋₆ alkyl, R^(c3) is C₁₋₆ alkyl optionallysubstituted by one substituent selected from the following Group A,R^(c4) is (i) hydrogen atom, or (ii) C₁₋₆ alkyl, and (6) a group offormula:

in which Y^(a) is a group selected from the following (i) to (iii): (i)C₁₋₆ alkylene, (ii) —C(═O)—, and (iii) —C(═O)—O—, Ring T is: (i) phenyl,(ii) C₃₋₆ cycloalkyl, or (iii) pyrrolidinyl, R^(c5) is (i) cyano, or(ii) nitro, p is an integer selected from 0 or 1, Group A is selectedfrom the group consisting of: (a) hydroxyl, (b) C₁₋₆ alkoxy, (c) cyano,(d) C₁₋₆ alkoxycarbonyl, (e) C₁₋₆ alkylcarbonyloxy, and (f) C₂₋₆alkenyloxy, or a pharmaceutically acceptable salt thereof, or a solvateof the compound or salt thereof.
 3. The compound as claimed in claim 1,wherein m1 is an integer of 0 or 1 and m2 is an integer of 1 or 2, or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 4. The compound as claimed in claim 3, wherein m1 is 1and m2 is 2, which is a compound of formula [II]:

wherein each symbol has the same meaning as defined in claim 1, or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 5. The compound as claimed in claim 3, wherein m1 is 0and m2 is 2, which is a compound of formula [III]:

wherein each symbol has the same meaning as defined in claim 1, or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 6. The compound as claimed in claim 3, wherein m1 is 0and m2 is 1, which is a compound of the general formula [IV]:

wherein each symbol has the same meaning as defined in claim 1, or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 7. The compound as claimed in claim 1, wherein m1 is 0and m2 is 3, or m1 is 2 and m2 is 1, or m1 is 2 and m2 is 2, or m1 is 3and m2 is 2, or a pharmaceutically acceptable salt thereof, or a solvateof the compound or salt thereof.
 8. (canceled)
 9. The compound asclaimed in claim 1, wherein n1 is 0 and n2 is 0, or a pharmaceuticallyacceptable salt thereof, or a solvate of the compound or salt thereof.10. The compound as claimed in claim 1, wherein n1 is 1 and n2 is 0, ora pharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 11. The compound as claimed in claim 1, wherein n1 is 0and n2 is 1, or a pharmaceutically acceptable salt thereof, or a solvateof the compound or salt thereof.
 12. The compound as claimed in claim 1,wherein n1 is 2 and n2 is 0, or a pharmaceutically acceptable saltthereof, or a solvate of the compound or salt thereof.
 13. The compoundas claimed in claim 1, wherein n1 is 0 and n2 is 2, or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 14. The compound as claimed in claim 10, wherein R^(a)is methyl or fluorine atom, or a pharmaceutically acceptable saltthereof, or a solvate of the compound or salt thereof.
 15. The compoundas claimed in claim 1, wherein R^(c) is —C(═O)—R^(c1), or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 16. The compound as claimed in claim 15, wherein R^(c1)is C₁₋₆ alkyl substituted by one hydroxyl or cyano group, or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof.
 17. The compound as claimed in claim 1, wherein R^(c)is —C(═O)—NR^(c3)R^(c4), or a pharmaceutically acceptable salt thereof,or a solvate of the compound or salt thereof.
 18. The compound asclaimed in claim 17, wherein R^(c3) is C₁₋₆ alkyl substituted by onecyano group, and R^(c4) is hydrogen, or a pharmaceutically acceptablesalt thereof, or a solvate of the compound or salt thereof. 19.-20.(canceled)
 21. A pharmaceutical composition, comprising the compound asclaimed in claim 1 or a pharmaceutically acceptable salt thereof, or asolvate of the compound or salt thereof, and a pharmaceuticallyacceptable carrier. 22.-27. (canceled)
 28. A method for inhibiting Januskinase 2 or Janus kinase 3, comprising administering to a mammal in needthereof a pharmaceutically effective amount of the compound as claimedin claim 1 or a pharmaceutically acceptable salt thereof, or a solvateof the compound or salt thereof.
 29. The method as claimed in claim 28,wherein the Janus kinase is Janus kinase
 3. 30. The method as claimed inclaim 28, wherein the Janus kinase is Janus kinase
 2. 31. A method fortreating or preventing a disease selected from the group consisting oforgan transplant rejection and graft versus host reaction aftertransplantation, comprising administering to a mammal in need thereof apharmaceutically effective amount of the compound as claimed in claim 1or a pharmaceutically acceptable salt thereof, or a solvate of thecompound or salt thereof.
 32. A method for treating rheumatoidarthritis, comprising administering to a mammal in need thereof apharmaceutically effective amount of the compound as claimed in claim 1or a pharmaceutically acceptable salt thereof, or a solvate of thecompound or salt thereof.
 33. A method for treating psoriasis,comprising administering to a mammal in need thereof a pharmaceuticallyeffective amount of the compound as claimed in claim 1 or apharmaceutically acceptable salt thereof, or a solvate of the compoundor salt thereof. 34.-39. (canceled)
 40. A method for treating atopicdermatitis, comprising administering to a mammal in need thereof apharmaceutically effective amount of the compound as claimed in claim 1or a pharmaceutically acceptable salt thereof, or a solvate of thecompound or salt thereof.
 41. A method for treating dry eye, comprisingadministering to a mammal in need thereof a pharmaceutically effectiveamount of the compound as claimed in claim 1 or a pharmaceuticallyacceptable salt thereof, or a solvate of the compound or salt thereof.